LOCUS       Exported                8904 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pWZL Neo Myr Flag PIM1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8904)
  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 8904)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8904
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     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(3369..3393)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3452..3913
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3508..3525)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3735..3754
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3918..4712
                     /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(3972..3991)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4582..4601
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             4899..5368
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        5404..5508
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5509..6369
                     /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(5727..5746)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8135..8723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8624..8643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8877..8894
                     /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 tgctcttgtc caaaatcaac tcgcttgccc
     2461 acctgcgcgc cgcgccctgc aacgacctgc acgccaccaa gctggcgccc ggcaaggaga
     2521 aggagcccct ggagtcgcag taccaggtgg gcccgctact gggcagcggc ggcttcggct
     2581 cggtctactc aggcatccgc gtctccgaca acttgccggt ggccatcaaa cacgtggaga
     2641 aggaccggat ttccgactgg ggagagctgc ctaatggcac tcgagtgccc atggaagtgg
     2701 tcctgctgaa gaaggtgagc tcgggtttct ccggcgtcat taggctcctg gactggttcg
     2761 agaggcccga cagtttcgtc ctgatcctgg agaggcccga gccggtgcaa gatctcttcg
     2821 acttcatcac ggaaagggga gccctgcaag aggagctggc ccgcagcttc ttctggcagg
     2881 tgctggaggc cgtgcggcac tgccacaact gcggggtgct ccaccgcgac atcaaggacg
     2941 aaaacatcct tatcgacctc aatcgcggcg agctcaagct catcgacttc gggtcggggg
     3001 cgctgctcaa ggacaccgtc tacacggact tcgatgggac ccgagtgtat agccctccag
     3061 agtggatccg ctaccatcgc taccatggca ggtcggcggc agtctggtcc ctggggatcc
     3121 tgctgtatga tatggtgtgt ggagatattc ctttcgagca tgacgaagag atcatcaggg
     3181 gccaggtttt cttcaggcag agggtctctt cagaatgtca gcatctcatt agatggtgct
     3241 tggccctgag accatcagat aggccaacct tcgaagaaat ccagaaccat ccatggatgc
     3301 aagatgttct cctgccccag gaaactgctg agatccacct ccacagcctg tcgccggggc
     3361 ccgcaaatac ccaactttct tgtacaaagt ggtttgatct agtcgaccaa ttccggttat
     3421 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     3481 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     3541 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
     3601 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     3661 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     3721 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     3781 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt
     3841 agtcgaggtt aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     3901 aacacgatga taataccatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3961 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4021 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4081 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4141 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4201 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4261 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4321 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4381 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4441 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4501 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4561 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4621 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4681 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg catcgataaa
     4741 ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt
     4801 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat
     4861 agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat
     4921 atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata
     4981 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     5041 tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg
     5101 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
     5161 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
     5221 tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc
     5281 tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga
     5341 ttgactaccc gtcagcgggg gtctttcaca ggccagtggg gccgcacttt tcggggaaat
     5401 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5461 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5521 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5581 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5641 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5701 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5761 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5821 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5881 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5941 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6001 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     6061 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6121 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6181 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6241 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6301 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6361 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt gcggccgcga
     6421 aaaggacaag cagcgaaaat tcacgccccc ttgggaggtg gcggcatatg caaaggatag
     6481 cactcccact ctactactgg gtatcatatg ctgactgtat atgcatgagg atagcatatg
     6541 ctacccggat acagattagg atagcatata ctacccagat atagattagg atagcatatg
     6601 ctacccagat atagattagg atagcctatg ctacccagat ataaattagg atagcatata
     6661 ctacccagat atagattagg atagcatatg ctacccagat atagattagg atagcctatg
     6721 ctacccagat atagattagg atagcatatg ctatccagat atttgggtag tatatgctac
     6781 ccatggcaac attagcccac cgtgctctca gcgacctcgt gaatatgagg accaacaacc
     6841 ctgtgcttgg cgctcaggcg caagtgtgtg taatttgtcc tccagatcgc agcaatcgcg
     6901 cccctatctt ggcccgccca cctacttatg caggtattcc ccggggtgcc attagtggtt
     6961 ttgtgggcaa gtggtttgac cgcagtggtt agcggggtta caatcagcca agttattaca
     7021 cccttatttt acagtccaaa accgcagggc ggcgtgtggg ggctgacgcg tgcccccact
     7081 ccacaatttc aaaaaaaaga gtggccactt gtctttgttt atgggcccca ttggcgtgga
     7141 gccccgttta attttcgggg gtgttagaga caaccagtgg agtccgctgc tgtcggcgtc
     7201 cactctcttt ccccttgtta caaatagagt gtaacaacat ggttcacctg tcttggtccc
     7261 tgcctgggac acatcttaat aaccccagta tcatattgca ctaggattat gtgttgccca
     7321 tagccataaa ttcgtgtgag atggacatcc agtctttacg gcttgtcccc accccatgga
     7381 tttctattgt taaagatatt cagaatgttt cattcctaca ctagtattta ttgcccaagg
     7441 ggtttgtgag ggttatattg gtgtcatagc acaatgccac cactgaaccc cccgtccaaa
     7501 ttttattctg ggggcgtcac ctgaaacctt gttttcgagc acctcacata caccttactg
     7561 ttcacaactc agcagttatt ctattagcta aacgaaggag aatgaagaag caggcgaaga
     7621 ttcaggagag ttcactgccc gctccttgat cttcagccac tgcccttgtg actaaaatgg
     7681 ttcactaccc tcgtggaatc ctgaccccat gtaaataaaa ccgtgacagc tcatggggtg
     7741 ggagatatcg ctgttcctta ggaccctttt actaacccta attcgatagc atatgcttcc
     7801 cgttgggtaa catatgctat tgaattaggg ttagtctgga tagtatatac tactacccgg
     7861 gaagcatatg ctacccgttt agggttaaca agggggcctt ataaacacta ttgctaatgc
     7921 cctcttgagg gtccgcttat cggtagctac acaggcccct ctgattgacg ttggtgtagc
     7981 ctcccgtagt cttcctgggc ccctgggagg tacatgtcgg ccgcaaaagg atctaggtga
     8041 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     8101 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     8161 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     8221 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     8281 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     8341 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     8401 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     8461 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     8521 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     8581 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     8641 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     8701 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     8761 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     8821 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     8881 agtcagtgag cgaggaagcg gaag
//
