LOCUS       Exported                9559 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pWZL Neo Myr Flag PKM2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9559)
  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 9559)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9559
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        69..448
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        449..652
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     603..623
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    873..1072
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1273..1689
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1619..1641
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1657..1673
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1707..1716
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1713..1754
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             1779..1802
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1809..1833
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3427..3451)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3510..3971
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3566..3583)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3793..3812
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3976..4770
                     /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(4030..4049)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4640..4659
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             4957..5426
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        5462..5566
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5567..6427
                     /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(5785..5804)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8193..8781
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8682..8701
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8935..8952
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9056..9483)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9166..9187)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9378..9397
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc
       61 gccatgttgg cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
      121 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      181 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      241 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      301 agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg
      361 gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat
      421 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg
      481 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      541 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc ccgccccgtt
      601 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
      661 gaaccgcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc
      721 tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga
      781 ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg ggagacccct
      841 gcccagggac caccgaccca ccaccgggag gcaagctggc cagcaactta tctgtgtctg
      901 tccgattgtc tagtgtctat gactgatttt atgcgcctgc gtcggtacta gttagctaac
      961 tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc cggccgcaac
     1021 cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg aggaagggag
     1081 tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga gaacctaaaa
     1141 cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg cgcgtcttgt
     1201 ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg
     1261 tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa
     1321 gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc
     1381 ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac
     1441 cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca
     1501 gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc
     1561 aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc
     1621 cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct
     1681 ctaggcgccg gccggatcca gaattcgcca ccatggggtc ttcaaaatct aaaccaaagg
     1741 accccagcca gcgccggcgc aggatccgag gttaccttga ctacaaagac gatgacgaca
     1801 agcaattgac aagtttgtac aaaaaagttg gcatgtcgaa gccccatagt gaagccggga
     1861 ctgccttcat tcagacccag cagctgcacg cagccatggc tgacacattc ctggagcaca
     1921 tgtgccgcct ggacattgat tcaccaccca tcacagcccg gaacactggc atcatctgta
     1981 ccattggccc agcttcccga tcagtggaga cgttgaagga gatgattaag tctggaatga
     2041 atgtggctcg tctgaacttc tctcatggaa ctcatgagta ccatgcggag accatcaaga
     2101 atgtgcgcac agccacggaa agctttgctt ctgaccccat cctctaccgg cccgttgctg
     2161 tggctctaga cactaaagga cctgagatcc gaactgggct catcaagggc agcggcactg
     2221 cagaggtgga gctgaagaag ggagccactc tcaaaatcac gctggataac gcctacatgg
     2281 aaaagtgtga cgagaacatc ctgtggctgg actacaagaa catctgcaag gtggtggaag
     2341 tgggcagcaa gatctacgtg gatgatgggc ttatttctct ccaggtgaag cagaaaggtg
     2401 ccgacttcct ggtgacggag gtggaaaatg gtggctcctt gggcagcaag aagggtgtga
     2461 accttcctgg ggctgctgtg gacttgcctg ctgtgtcgga gaaggacatc caggatctga
     2521 agtttggggt cgagcaggat gttgatatgg tgtttgcgtc attcatccgc aaggcatctg
     2581 atgtccatga agttaggaag gtcctgggag agaagggaaa gaacatcaag attatcagca
     2641 aaatcgagaa tcatgagggg gttcggaggt ttgatgaaat cctggaggcc agtgatggga
     2701 tcatggtggc tcgtggtgat ctaggcattg agattcctgc agagaaggtc ttccttgctc
     2761 agaagatgat gattggacgg tgcaaccgag ctgggaagcc tgtcatctgt gctactcaga
     2821 tgctggagag catgatcaag aagccccgcc ccactcgggc tgaaggcagt gatgtggcca
     2881 atgcagtcct ggatggagcc gactgcatca tgctgtctgg agaaacagcc aaaggggact
     2941 atcctctgga ggctgtgcgc atgcagcacc tgattgcccg tgaggcagag gctgccatct
     3001 accacttgca attatttgag gaactccgcc gcctggcgcc cattaccagc gaccccacag
     3061 aagccaccgc cgtgggtgcc gtggaggcct ccttcaagtg ctgcagtggg gccataatcg
     3121 tcctcaccaa gtctggcagg tctgctcacc aggtggccag ataccgccca cgtgccccca
     3181 tcattgctgt gacccggaat ccccagacag ctcgtcaggc ccacctgtac cgtggcatct
     3241 tccctgtgct gtgcaaggac ccagtccagg aggcctgggc tgaggacgtg gacctccggg
     3301 tgaactttgc catgaatgtt ggcaaggccc gaggcttctt caagaaggga gatgtggtca
     3361 ttgtgctgac cggatggcgc cctggctccg gcttcaccaa caccatgcgt gttgttcctg
     3421 tgccgttgcc aactttcttg tacaaagtgg tttgatctag tcgaccaatt ccggttattt
     3481 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     3541 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     3601 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     3661 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     3721 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     3781 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     3841 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     3901 tcgaggttaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     3961 cacgatgata ataccatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4021 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4081 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4141 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4201 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4261 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4321 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4381 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4441 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4501 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4561 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4621 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4681 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4741 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgca tcgataaaat
     4801 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     4861 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag
     4921 agaagttcag atcaaggtca ggaacagatg gaacagctga atatgggcca aacaggatat
     4981 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg
     5041 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     5101 gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt
     5161 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
     5221 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
     5281 ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc
     5341 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt
     5401 gactacccgt cagcgggggt ctttcacagg ccagtggggc cgcacttttc ggggaaatgt
     5461 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5521 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5581 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5641 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5701 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5761 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5821 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5881 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5941 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6001 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6061 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6121 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6181 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6241 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6301 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6361 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6421 ttggtaactg tcagaccaag tttactcata tatactttag attgatttgc ggccgcgaaa
     6481 aggacaagca gcgaaaattc acgccccctt gggaggtggc ggcatatgca aaggatagca
     6541 ctcccactct actactgggt atcatatgct gactgtatat gcatgaggat agcatatgct
     6601 acccggatac agattaggat agcatatact acccagatat agattaggat agcatatgct
     6661 acccagatat agattaggat agcctatgct acccagatat aaattaggat agcatatact
     6721 acccagatat agattaggat agcatatgct acccagatat agattaggat agcctatgct
     6781 acccagatat agattaggat agcatatgct atccagatat ttgggtagta tatgctaccc
     6841 atggcaacat tagcccaccg tgctctcagc gacctcgtga atatgaggac caacaaccct
     6901 gtgcttggcg ctcaggcgca agtgtgtgta atttgtcctc cagatcgcag caatcgcgcc
     6961 cctatcttgg cccgcccacc tacttatgca ggtattcccc ggggtgccat tagtggtttt
     7021 gtgggcaagt ggtttgaccg cagtggttag cggggttaca atcagccaag ttattacacc
     7081 cttattttac agtccaaaac cgcagggcgg cgtgtggggg ctgacgcgtg cccccactcc
     7141 acaatttcaa aaaaaagagt ggccacttgt ctttgtttat gggccccatt ggcgtggagc
     7201 cccgtttaat tttcgggggt gttagagaca accagtggag tccgctgctg tcggcgtcca
     7261 ctctctttcc ccttgttaca aatagagtgt aacaacatgg ttcacctgtc ttggtccctg
     7321 cctgggacac atcttaataa ccccagtatc atattgcact aggattatgt gttgcccata
     7381 gccataaatt cgtgtgagat ggacatccag tctttacggc ttgtccccac cccatggatt
     7441 tctattgtta aagatattca gaatgtttca ttcctacact agtatttatt gcccaagggg
     7501 tttgtgaggg ttatattggt gtcatagcac aatgccacca ctgaaccccc cgtccaaatt
     7561 ttattctggg ggcgtcacct gaaaccttgt tttcgagcac ctcacataca ccttactgtt
     7621 cacaactcag cagttattct attagctaaa cgaaggagaa tgaagaagca ggcgaagatt
     7681 caggagagtt cactgcccgc tccttgatct tcagccactg cccttgtgac taaaatggtt
     7741 cactaccctc gtggaatcct gaccccatgt aaataaaacc gtgacagctc atggggtggg
     7801 agatatcgct gttccttagg acccttttac taaccctaat tcgatagcat atgcttcccg
     7861 ttgggtaaca tatgctattg aattagggtt agtctggata gtatatacta ctacccggga
     7921 agcatatgct acccgtttag ggttaacaag ggggccttat aaacactatt gctaatgccc
     7981 tcttgagggt ccgcttatcg gtagctacac aggcccctct gattgacgtt ggtgtagcct
     8041 cccgtagtct tcctgggccc ctgggaggta catgtcggcc gcaaaaggat ctaggtgaag
     8101 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     8161 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     8221 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     8281 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     8341 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     8401 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     8461 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     8521 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     8581 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     8641 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     8701 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     8761 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     8821 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     8881 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     8941 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     9001 ccgattcatt aatgcaacta tggttggcgc gttgtaaacg ttaatatttt gttaaaattc
     9061 gcgttaaatt tttgttaaat cagctcattt tttaaccaat agaccgaaat cggcaaaatc
     9121 ccttataaat caaaagaata gcccgagata gagttgagtg ttgttccagt ttggaacaag
     9181 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     9241 gatggcccac tacgtgaacc atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa
     9301 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     9361 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctaaggc gctggcaagt
     9421 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     9481 gcgccgatct tcaatattgg ccattagcca tattattcat tggttatata gcataaatca
     9541 atattggcta ttggccatt
//
