LOCUS       sequence-157860-        8612 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 8612)
  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 8612)
  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 CDKN1B
FEATURES             Location/Qualifiers
     source          1..8612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(94..521)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(204..225)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     416..435
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     enhancer        666..1045
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1046..1249
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1200..1220
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    1470..1669
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1870..2286
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2216..2238
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2254..2270
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      2304..2313
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2310..2351
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             2376..2399
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    2406..2430
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3025..3049)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3108..3569
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3164..3181)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3391..3410
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3574..4368
                     /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(3628..3647)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4238..4257
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             4555..5024
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        5060..5164
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5165..5233,5234..6025)
                     /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(5383..5402)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7791..8379
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8280..8299
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8533..8550
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaactatg
       61 gttggcgcgt tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
      121 gctcattttt taaccaatag accgaaatcg gcaaaatccc ttataaatca aaagaatagc
      181 ccgagataga gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      241 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      301 cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      361 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      421 agaaagcgaa aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
      481 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gccgatcttc aatattggcc
      541 attagccata ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca
      601 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
      661 atgttggcat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      721 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      781 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      841 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      901 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
      961 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
     1021 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
     1081 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     1141 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac
     1201 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     1261 ccgcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct
     1321 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     1381 actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga gacccctgcc
     1441 cagggaccac cgacccacca ccgggaggca agctggccag caacttatct gtgtctgtcc
     1501 gattgtctag tgtctatgac tgattttatg cgcctgcgtc ggtactagtt agctaactag
     1561 ctctgtatct ggcggacccg tggtggaact gacgagttct gaacacccgg ccgcaaccct
     1621 gggagacgtc ccagggactt tgggggccgt ttttgtggcc cgacctgagg aagggagtcg
     1681 atgtggaatc cgaccccgtc aggatatgtg gttctggtag gagacgagaa cctaaaacag
     1741 ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc gaagccgcgc gtcttgtctg
     1801 ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct
     1861 gaaaattagg gccagactgt taccactccc ttaagtttga ccttaggtca ctggaaagat
     1921 gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga agagacgttg ggttaccttc
     1981 tgctctgcag aatggccaac ctttaacgtc ggatggccgc gagacggcac ctttaaccga
     2041 gacctcatca cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg acacccagac
     2101 caggtcccct acatcgtgac ctgggaagcc ttggcttttg acccccctcc ctgggtcaag
     2161 ccctttgtac accctaagcc tccgcctcct cttcctccat ccgccccgtc tctccccctt
     2221 gaacctcctc gttcgacccc gcctcgatcc tccctttatc cagccctcac tccttctcta
     2281 ggcgccggcc ggatccagaa ttcgccacca tggggtcttc aaaatctaaa ccaaaggacc
     2341 ccagccagcg ccggcgcagg atccgaggtt accttgacta caaagacgat gacgacaagc
     2401 aattgacaag tttgtacaaa aaagttggca tgtcaaacgt ccgagtgtct aacgggagcc
     2461 ctagcctgga gcggatggac gccaggcagg cggagcaccc caagccctcg gcctgcagga
     2521 acctcttcgg cccggtggac cacgaagagt taacccggga cttggagaag cactgcagag
     2581 acatggaaga ggcgagccag cgcaagtgga atttcgattt tcagaatcac aaacccctag
     2641 agggcaagta cgagtggcaa gaggtggaga agggcagctt gcccgagttc tactacagac
     2701 ccccgcggcc ccccaaaggt gcctgcaagg tgccggcgca ggagagccag gatggcagcg
     2761 ggagccgccc ggcggcgcct ttaattgggg ctccggctaa ctctgaggac acgcatttgg
     2821 tggacccaaa gactgatccg tcggacagcc agacggggtt agcggagcaa tgcgcaggaa
     2881 taaggaagcg acctgcaacc gacgattctt ctactcaaaa caaaagagcc aacagaacag
     2941 aagaaaatgt ttcagacggt tccccaaatg ccggttctgt ggagcagacg cccaagaagc
     3001 ctggcctcag aagacgtcaa acgtacccaa ctttcttgta caaagtggtt tgatctagtc
     3061 gaccaattcc ggttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     3121 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     3181 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     3241 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     3301 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     3361 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     3421 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     3481 tgctttacat gtgtttagtc gaggttaaaa aacgtctagg ccccccgaac cacggggacg
     3541 tggttttcct ttgaaaaaca cgatgataat accatgattg aacaagatgg attgcacgca
     3601 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3661 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3721 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     3781 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3841 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3901 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3961 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4021 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4081 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4141 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4201 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4261 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4321 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4381 ggttcgcatc gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag
     4441 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa
     4501 atacataact gagaatagag aagttcagat caaggtcagg aacagatgga acagctgaat
     4561 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     4621 atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     4681 cagggccaag aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc
     4741 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     4801 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     4861 gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg
     4921 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag
     4981 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcacaggcc agtggggccg
     5041 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5101 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5161 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5221 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5281 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5341 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5401 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5461 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5521 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5581 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5641 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5701 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5761 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5821 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5881 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5941 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6001 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6061 tgatttgcgg ccgcgaaaag gacaagcagc gaaaattcac gcccccttgg gaggtggcgg
     6121 catatgcaaa ggatagcact cccactctac tactgggtat catatgctga ctgtatatgc
     6181 atgaggatag catatgctac ccggatacag attaggatag catatactac ccagatatag
     6241 attaggatag catatgctac ccagatatag attaggatag cctatgctac ccagatataa
     6301 attaggatag catatactac ccagatatag attaggatag catatgctac ccagatatag
     6361 attaggatag cctatgctac ccagatatag attaggatag catatgctat ccagatattt
     6421 gggtagtata tgctacccat ggcaacatta gcccaccgtg ctctcagcga cctcgtgaat
     6481 atgaggacca acaaccctgt gcttggcgct caggcgcaag tgtgtgtaat ttgtcctcca
     6541 gatcgcagca atcgcgcccc tatcttggcc cgcccaccta cttatgcagg tattccccgg
     6601 ggtgccatta gtggttttgt gggcaagtgg tttgaccgca gtggttagcg gggttacaat
     6661 cagccaagtt attacaccct tattttacag tccaaaaccg cagggcggcg tgtgggggct
     6721 gacgcgtgcc cccactccac aatttcaaaa aaaagagtgg ccacttgtct ttgtttatgg
     6781 gccccattgg cgtggagccc cgtttaattt tcgggggtgt tagagacaac cagtggagtc
     6841 cgctgctgtc ggcgtccact ctctttcccc ttgttacaaa tagagtgtaa caacatggtt
     6901 cacctgtctt ggtccctgcc tgggacacat cttaataacc ccagtatcat attgcactag
     6961 gattatgtgt tgcccatagc cataaattcg tgtgagatgg acatccagtc tttacggctt
     7021 gtccccaccc catggatttc tattgttaaa gatattcaga atgtttcatt cctacactag
     7081 tatttattgc ccaaggggtt tgtgagggtt atattggtgt catagcacaa tgccaccact
     7141 gaaccccccg tccaaatttt attctggggg cgtcacctga aaccttgttt tcgagcacct
     7201 cacatacacc ttactgttca caactcagca gttattctat tagctaaacg aaggagaatg
     7261 aagaagcagg cgaagattca ggagagttca ctgcccgctc cttgatcttc agccactgcc
     7321 cttgtgacta aaatggttca ctaccctcgt ggaatcctga ccccatgtaa ataaaaccgt
     7381 gacagctcat ggggtgggag atatcgctgt tccttaggac ccttttacta accctaattc
     7441 gatagcatat gcttcccgtt gggtaacata tgctattgaa ttagggttag tctggatagt
     7501 atatactact acccgggaag catatgctac ccgtttaggg ttaacaaggg ggccttataa
     7561 acactattgc taatgccctc ttgagggtcc gcttatcggt agctacacag gcccctctga
     7621 ttgacgttgg tgtagcctcc cgtagtcttc ctgggcccct gggaggtaca tgtcggccgc
     7681 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     7741 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     7801 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     7861 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     7921 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7981 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     8041 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     8101 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     8161 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     8221 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     8281 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     8341 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     8401 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     8461 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     8521 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     8581 cctctccccg cgcgttggcc gattcattaa tg
//
