LOCUS       sequence-442157-        8668 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8668)
  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 8668)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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 TK1
FEATURES             Location/Qualifiers
     source          1..8668
                     /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(2616..2640)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2699..3160
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2755..2772)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2982..3001
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3165..3959
                     /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(3219..3238)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3829..3848
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             4146..4615
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        4651..4755
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4756..4824,4825..5616)
                     /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(4974..4993)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7382..7970
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7871..7890
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8124..8141
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(8245..8668,1..4))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8355..8376)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8567..8586
                     /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 gcatgagctg
     1921 cattaacctg cccactgtgc tgcccggctc ccccagcaag acccgggggc agatccaggt
     1981 gattctcggg ccgatgttct caggaaaaag cacagagttg atgagacgcg tccgtcgctt
     2041 ccagattgct cagtacaagt gcctggtgat caagtatgcc aaagacactc gctacagcag
     2101 cagcttctgc acacatgacc ggaacaccat ggaggcgctg cccgcctgcc tgctccgaga
     2161 cgtggcccag gaggccctgg gcgtggctgt cataggcatc gacgaggggc agtttttccc
     2221 tgacatcgtg gagttctgcg aggccatggc caacgccggg aagaccgtaa ttgtggctgc
     2281 actggatggg accttccaga ggaagccatt tggggccatc ctgaacctgg tgccgctggc
     2341 cgagagcgtg gtgaagctga cggcggtgtg catggagtgc ttccgggaag ccgcctatac
     2401 caagaggctc ggcacagaga aggaggtcga ggtgattggg ggagcagaca agtaccactc
     2461 cgtgtgtcgg ctctgctact tcaagaaggc ctcaggccag cctgccgggc cggacaacaa
     2521 agagaactgc ccagtgccag gaaagccagg ggaagccgtg gctgccagga agctctttgc
     2581 cccacagcag attctgcaat gcagccctgc caactgccca actttcttgt acaaagtggt
     2641 ttgatctagt cgaccaattc cggttatttt ccaccatatt gccgtctttt ggcaatgtga
     2701 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     2761 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     2821 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     2881 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     2941 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     3001 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     3061 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaacgtctag gccccccgaa
     3121 ccacggggac gtggttttcc tttgaaaaac acgatgataa taccatgatt gaacaagatg
     3181 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3241 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3301 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3361 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3421 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3481 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3541 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3601 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3661 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     3721 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3781 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     3841 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3901 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     3961 cgggactctg gggttcgcat cgataaaata aaagatttta tttagtctcc agaaaaaggg
     4021 gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa
     4081 ggcatggaaa aatacataac tgagaataga gaagttcaga tcaaggtcag gaacagatgg
     4141 aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     4201 gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
     4261 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt
     4321 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
     4381 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
     4441 tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc
     4501 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
     4561 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcacaggc
     4621 cagtggggcc gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4681 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4741 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4801 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4861 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4921 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4981 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5041 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5101 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5161 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5221 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5281 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5341 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5401 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5461 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5521 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5581 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5641 atactttaga ttgatttgcg gccgcgaaaa ggacaagcag cgaaaattca cgcccccttg
     5701 ggaggtggcg gcatatgcaa aggatagcac tcccactcta ctactgggta tcatatgctg
     5761 actgtatatg catgaggata gcatatgcta cccggataca gattaggata gcatatacta
     5821 cccagatata gattaggata gcatatgcta cccagatata gattaggata gcctatgcta
     5881 cccagatata aattaggata gcatatacta cccagatata gattaggata gcatatgcta
     5941 cccagatata gattaggata gcctatgcta cccagatata gattaggata gcatatgcta
     6001 tccagatatt tgggtagtat atgctaccca tggcaacatt agcccaccgt gctctcagcg
     6061 acctcgtgaa tatgaggacc aacaaccctg tgcttggcgc tcaggcgcaa gtgtgtgtaa
     6121 tttgtcctcc agatcgcagc aatcgcgccc ctatcttggc ccgcccacct acttatgcag
     6181 gtattccccg gggtgccatt agtggttttg tgggcaagtg gtttgaccgc agtggttagc
     6241 ggggttacaa tcagccaagt tattacaccc ttattttaca gtccaaaacc gcagggcggc
     6301 gtgtgggggc tgacgcgtgc ccccactcca caatttcaaa aaaaagagtg gccacttgtc
     6361 tttgtttatg ggccccattg gcgtggagcc ccgtttaatt ttcgggggtg ttagagacaa
     6421 ccagtggagt ccgctgctgt cggcgtccac tctctttccc cttgttacaa atagagtgta
     6481 acaacatggt tcacctgtct tggtccctgc ctgggacaca tcttaataac cccagtatca
     6541 tattgcacta ggattatgtg ttgcccatag ccataaattc gtgtgagatg gacatccagt
     6601 ctttacggct tgtccccacc ccatggattt ctattgttaa agatattcag aatgtttcat
     6661 tcctacacta gtatttattg cccaaggggt ttgtgagggt tatattggtg tcatagcaca
     6721 atgccaccac tgaacccccc gtccaaattt tattctgggg gcgtcacctg aaaccttgtt
     6781 ttcgagcacc tcacatacac cttactgttc acaactcagc agttattcta ttagctaaac
     6841 gaaggagaat gaagaagcag gcgaagattc aggagagttc actgcccgct ccttgatctt
     6901 cagccactgc ccttgtgact aaaatggttc actaccctcg tggaatcctg accccatgta
     6961 aataaaaccg tgacagctca tggggtggga gatatcgctg ttccttagga cccttttact
     7021 aaccctaatt cgatagcata tgcttcccgt tgggtaacat atgctattga attagggtta
     7081 gtctggatag tatatactac tacccgggaa gcatatgcta cccgtttagg gttaacaagg
     7141 gggccttata aacactattg ctaatgccct cttgagggtc cgcttatcgg tagctacaca
     7201 ggcccctctg attgacgttg gtgtagcctc ccgtagtctt cctgggcccc tgggaggtac
     7261 atgtcggccg caaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7321 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7381 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7441 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7501 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     7561 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7621 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7681 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7741 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7801 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7861 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7921 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7981 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     8041 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     8101 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     8161 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcaactat ggttggcgcg
     8221 ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     8281 ttaaccaata gaccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag
     8341 agttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     8401 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat
     8461 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     8521 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     8581 aaggagcggg cgctaaggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     8641 ccgccgcgct taatgcgccg ctacaggg
//
