LOCUS       sequence-172643-        8796 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 8796)
  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 8796)
  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 LIMK1
FEATURES             Location/Qualifiers
     source          1..8796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        58..437
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        438..641
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     592..612
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    862..1061
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1262..1678
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1608..1630
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1646..1662
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1696..1705
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1702..1743
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             1768..1791
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1798..1822
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2653..2677)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2736..3197
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2792..2809)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3019..3038
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3202..3996
                     /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(3256..3275)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3866..3885
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             4183..4652
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        4688..4792
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4793..4861,4862..5653)
                     /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(5011..5030)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7419..8007
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7908..7927
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8161..8178
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8282..8709)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8392..8413)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8604..8623
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttggc
       61 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      121 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      181 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      241 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      301 tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      361 attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag
      421 tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt
      481 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      541 accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg
      601 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgcgcca
      661 gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg
      721 catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg
      781 tcagcggggg tctttcattt gggggctcgt ccgggatcgg gagacccctg cccagggacc
      841 accgacccac caccgggagg caagctggcc agcaacttat ctgtgtctgt ccgattgtct
      901 agtgtctatg actgatttta tgcgcctgcg tcggtactag ttagctaact agctctgtat
      961 ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg
     1021 tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt cgatgtggaa
     1081 tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc
     1141 tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg
     1201 ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta
     1261 gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg
     1321 gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc
     1381 agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat
     1441 cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc
     1501 ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt
     1561 acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc
     1621 tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc taggcgccgg
     1681 ccggatccag aattcgccac catggggtct tcaaaatcta aaccaaagga ccccagccag
     1741 cgccggcgca ggatccgagg ttaccttgac tacaaagacg atgacgacaa gcaattgaca
     1801 agtttgtaca aaaaagttgg catgaggttg acgctacttt gttgcacctg gagggaagaa
     1861 cgtatgggag aggaaggaag cgagttgccc gtgtgtgcaa gctgcggcca gaggatctat
     1921 gatggccagt acctccaggc cctgaacgcg gactggcacg cagactgctt caggtgttgt
     1981 gactgcagtg cctccctgtc gcaccagtac tatgagaagg atgggcagct cttctgcaag
     2041 aaggactact gggcccgcta tggcgagtcc tgccatgggt gctctgagca aatcaccaag
     2101 ggactggtta tggtggctgg ggagctgaag taccaccccg agtgtttcat ctgcctcacg
     2161 tgtgggacct ttatcggtga cggggacacc tacacgctgg tggagcactc caagctgtac
     2221 tgcgggcact gctactacca gactgtggtg acccccgtca tcgagcagat cctgcctgac
     2281 tcccctggct cccacctgcc ccacaccgtc accctggtgt ccatcccagc ctcatctcat
     2341 ggcaagcgtg gactttcagt ctccattgac cccccgcacg gcccaccggg ctgtggcacc
     2401 gagcactcac acaccgtccg cgtccaggga gtggatccgg gctgcatgag cccagatgtg
     2461 aagaattcca tccacgtcgg agaccggatc ttggaaatca atggcacgcc catccgaaat
     2521 gtgcccctgg acgagattga cctgctgatt caggaaacca gccgcctgct ccagctgacc
     2581 ctcgagcatg accctcacga tacactgggc cacgggctgg ggcctgagac cagccccctg
     2641 aggtccccga cttgccaact ttcttgtaca aagtggtttg atctagtcga ccaattccgg
     2701 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     2761 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     2821 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     2881 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     2941 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     3001 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     3061 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     3121 gtttagtcga ggttaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     3181 gaaaaacacg atgataatac catgattgaa caagatggat tgcacgcagg ttctccggcc
     3241 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3301 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3361 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     3421 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3481 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3541 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3601 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3661 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3721 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3781 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3841 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3901 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     3961 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcatcga
     4021 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     4081 ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga
     4141 gaatagagaa gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca
     4201 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg
     4261 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     4321 cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc
     4381 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     4441 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     4501 tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa
     4561 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     4621 gtgattgact acccgtcagc gggggtcttt cacaggccag tggggccgca cttttcgggg
     4681 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4741 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4801 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4861 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4921 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4981 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5041 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5101 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5161 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5221 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5281 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5341 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5401 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5461 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5521 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5581 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5641 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttgcggcc
     5701 gcgaaaagga caagcagcga aaattcacgc ccccttggga ggtggcggca tatgcaaagg
     5761 atagcactcc cactctacta ctgggtatca tatgctgact gtatatgcat gaggatagca
     5821 tatgctaccc ggatacagat taggatagca tatactaccc agatatagat taggatagca
     5881 tatgctaccc agatatagat taggatagcc tatgctaccc agatataaat taggatagca
     5941 tatactaccc agatatagat taggatagca tatgctaccc agatatagat taggatagcc
     6001 tatgctaccc agatatagat taggatagca tatgctatcc agatatttgg gtagtatatg
     6061 ctacccatgg caacattagc ccaccgtgct ctcagcgacc tcgtgaatat gaggaccaac
     6121 aaccctgtgc ttggcgctca ggcgcaagtg tgtgtaattt gtcctccaga tcgcagcaat
     6181 cgcgccccta tcttggcccg cccacctact tatgcaggta ttccccgggg tgccattagt
     6241 ggttttgtgg gcaagtggtt tgaccgcagt ggttagcggg gttacaatca gccaagttat
     6301 tacaccctta ttttacagtc caaaaccgca gggcggcgtg tgggggctga cgcgtgcccc
     6361 cactccacaa tttcaaaaaa aagagtggcc acttgtcttt gtttatgggc cccattggcg
     6421 tggagccccg tttaattttc gggggtgtta gagacaacca gtggagtccg ctgctgtcgg
     6481 cgtccactct ctttcccctt gttacaaata gagtgtaaca acatggttca cctgtcttgg
     6541 tccctgcctg ggacacatct taataacccc agtatcatat tgcactagga ttatgtgttg
     6601 cccatagcca taaattcgtg tgagatggac atccagtctt tacggcttgt ccccacccca
     6661 tggatttcta ttgttaaaga tattcagaat gtttcattcc tacactagta tttattgccc
     6721 aaggggtttg tgagggttat attggtgtca tagcacaatg ccaccactga accccccgtc
     6781 caaattttat tctgggggcg tcacctgaaa ccttgttttc gagcacctca catacacctt
     6841 actgttcaca actcagcagt tattctatta gctaaacgaa ggagaatgaa gaagcaggcg
     6901 aagattcagg agagttcact gcccgctcct tgatcttcag ccactgccct tgtgactaaa
     6961 atggttcact accctcgtgg aatcctgacc ccatgtaaat aaaaccgtga cagctcatgg
     7021 ggtgggagat atcgctgttc cttaggaccc ttttactaac cctaattcga tagcatatgc
     7081 ttcccgttgg gtaacatatg ctattgaatt agggttagtc tggatagtat atactactac
     7141 ccgggaagca tatgctaccc gtttagggtt aacaaggggg ccttataaac actattgcta
     7201 atgccctctt gagggtccgc ttatcggtag ctacacaggc ccctctgatt gacgttggtg
     7261 tagcctcccg tagtcttcct gggcccctgg gaggtacatg tcggccgcaa aaggatctag
     7321 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7381 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7441 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7501 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7561 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7621 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7681 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7741 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7801 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7861 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7921 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7981 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     8041 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     8101 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     8161 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     8221 cgttggccga ttcattaatg caactatggt tggcgcgttg taaacgttaa tattttgtta
     8281 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaatagac cgaaatcggc
     8341 aaaatccctt ataaatcaaa agaatagccc gagatagagt tgagtgttgt tccagtttgg
     8401 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     8461 cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc
     8521 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     8581 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc taaggcgctg
     8641 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     8701 cagggcgcgc cgatcttcaa tattggccat tagccatatt attcattggt tatatagcat
     8761 aaatcaatat tggctattgg ccattgcata cgttgt
//
