LOCUS       sequence-203982-        8692 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 8692)
  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 8692)
  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 AKAP8L
FEATURES             Location/Qualifiers
     source          1..8692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    805..1004
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1205..1621
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1551..1573
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1589..1605
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1639..1648
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1645..1686
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             1711..1734
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1741..1765
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2492..2516)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2575..3036
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2631..2648)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2858..2877
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3041..3835
                     /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(3095..3114)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3705..3724
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             4022..4491
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        4527..4631
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4632..4700,4701..5492)
                     /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(4850..4869)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7258..7846
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7747..7766
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8000..8017
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8121..8548)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8231..8252)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8443..8462
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgcg
      601 ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag
      661 ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac
      721 ccgtcagcgg gggtctttca tttgggggct cgtccgggat cgggagaccc ctgcccaggg
      781 accaccgacc caccaccggg aggcaagctg gccagcaact tatctgtgtc tgtccgattg
      841 tctagtgtct atgactgatt ttatgcgcct gcgtcggtac tagttagcta actagctctg
      901 tatctggcgg acccgtggtg gaactgacga gttctgaaca cccggccgca accctgggag
      961 acgtcccagg gactttgggg gccgtttttg tggcccgacc tgaggaaggg agtcgatgtg
     1021 gaatccgacc ccgtcaggat atgtggttct ggtaggagac gagaacctaa aacagttccc
     1081 gcctccgtct gaatttttgc tttcggtttg gaaccgaagc cgcgcgtctt gtctgctgca
     1141 gcgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa
     1201 ttagggccag actgttacca ctcccttaag tttgacctta ggtcactgga aagatgtcga
     1261 gcggatcgct cacaaccagt cggtagatgt caagaagaga cgttgggtta ccttctgctc
     1321 tgcagaatgg ccaaccttta acgtcggatg gccgcgagac ggcaccttta accgagacct
     1381 catcacccag gttaagatca aggtcttttc acctggcccg catggacacc cagaccaggt
     1441 cccctacatc gtgacctggg aagccttggc ttttgacccc cctccctggg tcaagccctt
     1501 tgtacaccct aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc
     1561 tcctcgttcg accccgcctc gatcctccct ttatccagcc ctcactcctt ctctaggcgc
     1621 cggccggatc cagaattcgc caccatgggg tcttcaaaat ctaaaccaaa ggaccccagc
     1681 cagcgccggc gcaggatccg aggttacctt gactacaaag acgatgacga caagcaattg
     1741 acaagtttgt acaaaaaagt tggcatgagc tacacaggct ttgtccaggg atctgaaacc
     1801 actttgcagt cgacatactc ggataccagc gctcagccca cctgtgatta tggatatgga
     1861 acttggaact ctgggacaaa tagaggctac gagggctatg gctatggcta tggctatggc
     1921 caggataaca ccaccaacta tgggtatggt atggccactt cacactcttg ggaaatgcct
     1981 agctctgaca caaatgcaaa cactagtgcc tcgggtagcg ccagtgccga ttccgtttta
     2041 tccagaatta accagcgctt agatatggtg ccgcatttgg agacagacat gatgcaagga
     2101 ggcgtgtacg gctcaggtgg agaaaggtat gactcttatg agtcctgcga ctcgagggcc
     2161 gtcctgagtg agcgcgacct gtaccggtca ggctatgact acagcgagct tgaccctgag
     2221 atggaaatgg cctatgaggg ccaatacgat gcctaccgcg accagttccg catgcgtggc
     2281 aacgacacct tcggtcccag ggcacagggc tgggcccggg atgcccggag cggccggcca
     2341 atggcctcag gctatgggcg catgtgggaa gaccccatgg gggcccgggg ccagtgcatg
     2401 tctggtgcct ctcggctgcc ctccctcttc tcccagaaca tcatccccga gtacggcatg
     2461 ttccagggca tgcgaggcgg gggcgccccg tgcccaactt tcttgtacaa agtggtttga
     2521 tctagtcgac caattccggt tattttccac catattgccg tcttttggca atgtgagggc
     2581 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     2641 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     2701 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     2761 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     2821 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     2881 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     2941 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaac gtctaggccc cccgaaccac
     3001 ggggacgtgg ttttcctttg aaaaacacga tgataatacc atgattgaac aagatggatt
     3061 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3121 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3181 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3241 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3301 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3361 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3421 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3481 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3541 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     3601 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3661 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3721 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3781 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     3841 actctggggt tcgcatcgat aaaataaaag attttattta gtctccagaa aaagggggga
     3901 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
     3961 tggaaaaata cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca
     4021 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     4081 agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     4141 cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta
     4201 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     4261 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     4321 taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg
     4381 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
     4441 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc acaggccagt
     4501 ggggccgcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4561 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4621 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4681 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4741 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4801 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4861 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4921 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4981 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5041 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5101 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5161 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5221 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5281 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5341 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5401 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5461 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5521 tttagattga tttgcggccg cgaaaaggac aagcagcgaa aattcacgcc cccttgggag
     5581 gtggcggcat atgcaaagga tagcactccc actctactac tgggtatcat atgctgactg
     5641 tatatgcatg aggatagcat atgctacccg gatacagatt aggatagcat atactaccca
     5701 gatatagatt aggatagcat atgctaccca gatatagatt aggatagcct atgctaccca
     5761 gatataaatt aggatagcat atactaccca gatatagatt aggatagcat atgctaccca
     5821 gatatagatt aggatagcct atgctaccca gatatagatt aggatagcat atgctatcca
     5881 gatatttggg tagtatatgc tacccatggc aacattagcc caccgtgctc tcagcgacct
     5941 cgtgaatatg aggaccaaca accctgtgct tggcgctcag gcgcaagtgt gtgtaatttg
     6001 tcctccagat cgcagcaatc gcgcccctat cttggcccgc ccacctactt atgcaggtat
     6061 tccccggggt gccattagtg gttttgtggg caagtggttt gaccgcagtg gttagcgggg
     6121 ttacaatcag ccaagttatt acacccttat tttacagtcc aaaaccgcag ggcggcgtgt
     6181 gggggctgac gcgtgccccc actccacaat ttcaaaaaaa agagtggcca cttgtctttg
     6241 tttatgggcc ccattggcgt ggagccccgt ttaattttcg ggggtgttag agacaaccag
     6301 tggagtccgc tgctgtcggc gtccactctc tttccccttg ttacaaatag agtgtaacaa
     6361 catggttcac ctgtcttggt ccctgcctgg gacacatctt aataacccca gtatcatatt
     6421 gcactaggat tatgtgttgc ccatagccat aaattcgtgt gagatggaca tccagtcttt
     6481 acggcttgtc cccaccccat ggatttctat tgttaaagat attcagaatg tttcattcct
     6541 acactagtat ttattgccca aggggtttgt gagggttata ttggtgtcat agcacaatgc
     6601 caccactgaa ccccccgtcc aaattttatt ctgggggcgt cacctgaaac cttgttttcg
     6661 agcacctcac atacacctta ctgttcacaa ctcagcagtt attctattag ctaaacgaag
     6721 gagaatgaag aagcaggcga agattcagga gagttcactg cccgctcctt gatcttcagc
     6781 cactgccctt gtgactaaaa tggttcacta ccctcgtgga atcctgaccc catgtaaata
     6841 aaaccgtgac agctcatggg gtgggagata tcgctgttcc ttaggaccct tttactaacc
     6901 ctaattcgat agcatatgct tcccgttggg taacatatgc tattgaatta gggttagtct
     6961 ggatagtata tactactacc cgggaagcat atgctacccg tttagggtta acaagggggc
     7021 cttataaaca ctattgctaa tgccctcttg agggtccgct tatcggtagc tacacaggcc
     7081 cctctgattg acgttggtgt agcctcccgt agtcttcctg ggcccctggg aggtacatgt
     7141 cggccgcaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7201 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     7261 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     7321 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     7381 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     7441 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7501 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7561 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7621 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7681 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7741 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7801 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7861 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     7921 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     7981 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     8041 caaaccgcct ctccccgcgc gttggccgat tcattaatgc aactatggtt ggcgcgttgt
     8101 aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     8161 ccaatagacc gaaatcggca aaatccctta taaatcaaaa gaatagcccg agatagagtt
     8221 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     8281 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac ccaaatcaag
     8341 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     8401 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     8461 agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     8521 cgcgcttaat gcgccgctac agggcgcgcc gatcttcaat attggccatt agccatatta
     8581 ttcattggtt atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta
     8641 tatcataata tgtacattta tattggctca tgtccaatat gaccgccatg tt
//
