LOCUS       Exported                6631 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pBabe-Puro-Myr-Flag-AKT1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6631)
  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.
  JOURNAL   Cell. 2007 Jun 15. 129(6):1065-1079.
  PUBMED    17574021
REFERENCE   2  (bases 1 to 6631)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6631
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(982..1842)
                     /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     1605..1624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1843..1947)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             1978..2447
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    2512..2711
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             2912..3328
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3258..3280
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3296..3312
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             3339..3380
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             3405..3428
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    3435..3459
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3459..4895
                     /codon_start=1
                     /gene="AKT1"
                     /product="human serine/threonine protein kinase activated 
                     by growth factors"
                     /label=Akt1
                     /translation="MSDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDV
                     DQREAPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWTTAIQ
                     TVADGLKKQEEEEMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGK
                     VILVKEKATGRYYAMKILKKEVIVAKDEVAHTLTENRVLQNSRHPFLTALKYSFQTHDR
                     LCFVMEYANGGELFFHLSRERVFSEDRARFYGAEIVSALDYLHSEKNVVYRDLKLENLM
                     LDKDGHIKITDFGLCKEGIKDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMY
                     EMMCGRLPFYNQDHEKLFELILMEEIRFPRTLGPEAKSLLSGLLKKDPKQRLGGGSEDA
                     KEIMQHRFFAGIVWQHVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITITPPDQDDSM
                     ECVDSERRPHFPQFSYSASGT"
     primer_bind     complement(4931..4951)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4936..5265
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5116..5251
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5178..5197
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5275..5874
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5275..5294)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5771..5791
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6095..6564
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttgcggccgc aaatcaatct aaagtatata
      961 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1021 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1081 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     1141 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     1201 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     1261 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     1321 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     1381 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1441 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     1501 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     1561 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     1621 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1681 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1741 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     1801 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     1861 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     1921 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcagcctg
     1981 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     2041 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     2101 gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga
     2161 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     2221 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     2281 agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac
     2341 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     2401 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt
     2461 ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg caagctggcc
     2521 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg
     2581 tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
     2641 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
     2701 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
     2761 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
     2821 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
     2881 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
     2941 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
     3001 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
     3061 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
     3121 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
     3181 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
     3241 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
     3301 tccagccctc actccttctc taggcgccgg ccggatctat ggggtcttca aaatctaaac
     3361 caaaggaccc cagccagcgc cggcgcagga tccgaggtta ccttgactac aaagacgatg
     3421 acgacaagca attgacaagt ttgtacaaaa aagttggcat gagcgacgtg gctattgtga
     3481 aggagggttg gctgcacaaa cgaggggagt acatcaagac ctggcggcca cgctacttcc
     3541 tcctcaagaa tgatggcacc ttcattggct acaaggagcg gccgcaggat gtggaccaac
     3601 gtgaggctcc cctcaacaac ttctctgtgg cgcagtgcca gctgatgaag acggagcggc
     3661 cccggcccaa caccttcatc atccgctgcc tgcagtggac cactgtcatc gaacgcacct
     3721 tccatgtgga gactcctgag gagcgggagg agtggacaac cgccatccag actgtggctg
     3781 acggcctcaa gaagcaggag gaggaggaga tggacttccg gtcgggctca cccagtgaca
     3841 actcaggggc tgaagagatg gaggtgtccc tggccaagcc caagcaccgc gtgaccatga
     3901 acgagtttga gtacctgaag ctgctgggca agggcacttt cggcaaggtg atcctggtga
     3961 aggagaaggc cacaggccgc tactacgcca tgaagatcct caagaaggaa gtcatcgtgg
     4021 ccaaggacga ggtggcccac acactcaccg agaaccgcgt cctgcagaac tccaggcacc
     4081 ccttcctcac agccctgaag tactctttcc agacccacga ccgcctctgc tttgtcatgg
     4141 agtacgccaa cgggggcgag ctgttcttcc acctgtcccg ggagcgtgtg ttctccgagg
     4201 accgggcccg cttctatggc gctgagattg tgtcagccct ggactacctg cactcggaga
     4261 agaacgtggt gtaccgggac ctcaagctgg agaacctcat gctggacaag gacgggcaca
     4321 ttaagatcac agacttcggg ctgtgcaagg aggggatcaa ggacggtgcc accatgaaga
     4381 ccttttgcgg cacacctgag tacctggccc ccgaggtgct ggaggacaat gactacggcc
     4441 gtgcagtgga ctggtggggg ctgggcgtgg tcatgtacga gatgatgtgc ggtcgcctgc
     4501 ccttctacaa ccaggaccat gagaagcttt ttgagctcat cctcatggag gagatccgct
     4561 tcccgcgcac gcttggtccc gaggccaagt ccttgctttc agggctgctc aagaaggacc
     4621 ccaagcagag gcttggcggg ggctccgagg acgccaagga gatcatgcag catcgcttct
     4681 ttgccggtat cgtgtggcag cacgtgtacg agaagaagct cagcccaccc ttcaagcccc
     4741 aggtcacgtc ggagactgac accaggtatt ttgatgagga gttcacggcc cagatgatca
     4801 ccatcacacc acctgaccaa gatgacagca tggagtgtgt ggacagcgag cgcaggcccc
     4861 acttccccca gttctcctac tcggccagcg gcacggcgta ggaattcgcc agcacagtgg
     4921 tcgaccctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4981 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     5041 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     5101 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     5161 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     5221 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct taccatgacc
     5281 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     5341 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     5401 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     5461 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     5521 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     5581 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     5641 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     5701 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     5761 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     5821 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg
     5881 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc gataaaataa
     5941 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     6001 aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag
     6061 aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct
     6121 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg
     6181 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     6241 ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     6301 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     6361 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     6421 ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt
     6481 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     6541 ctacccgtca gcgggggtct ttcacatgca gcatgtatca aaattaattt ggtttttttt
     6601 cttaagtatt tacattaaat ggccatagtt g
//
