LOCUS       Exported                8133 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pWZL Neo Myr Flag STK4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8133)
  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 8133)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8133
                     /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(2546..2570)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2629..3090
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2685..2702)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2912..2931
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3095..3889
                     /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(3149..3168)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3759..3778
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             4076..4545
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        4581..4685
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4686..5546
                     /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(4904..4923)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7312..7900
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7801..7820
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8054..8071
                     /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 tggagacggt acagctgagg aacccgccgc
     2461 gccggcagct gaaaaagttg gatgaagata gtttaaccaa acaaccagaa gaagtatttg
     2521 atgtctagag aacttggaga agggtgccca actttcttgt acaaagtggt ttgatctagt
     2581 cgaccaattc cggttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     2641 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     2701 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     2761 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     2821 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     2881 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     2941 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     3001 atgctttaca tgtgtttagt cgaggttaaa aaacgtctag gccccccgaa ccacggggac
     3061 gtggttttcc tttgaaaaac acgatgataa taccatgatt gaacaagatg gattgcacgc
     3121 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3181 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     3241 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     3301 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     3361 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     3421 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     3481 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     3541 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     3601 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     3661 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     3721 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     3781 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     3841 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     3901 gggttcgcat cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa
     3961 gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa
     4021 aatacataac tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa
     4081 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     4141 gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     4201 tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac
     4261 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
     4321 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
     4381 agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc
     4441 gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga
     4501 gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcacaggc cagtggggcc
     4561 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4621 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4681 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4741 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4801 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4861 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4921 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4981 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5041 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5101 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5161 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5221 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5281 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5341 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5401 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5461 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5521 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5581 ttgatttgcg gccgcgaaaa ggacaagcag cgaaaattca cgcccccttg ggaggtggcg
     5641 gcatatgcaa aggatagcac tcccactcta ctactgggta tcatatgctg actgtatatg
     5701 catgaggata gcatatgcta cccggataca gattaggata gcatatacta cccagatata
     5761 gattaggata gcatatgcta cccagatata gattaggata gcctatgcta cccagatata
     5821 aattaggata gcatatacta cccagatata gattaggata gcatatgcta cccagatata
     5881 gattaggata gcctatgcta cccagatata gattaggata gcatatgcta tccagatatt
     5941 tgggtagtat atgctaccca tggcaacatt agcccaccgt gctctcagcg acctcgtgaa
     6001 tatgaggacc aacaaccctg tgcttggcgc tcaggcgcaa gtgtgtgtaa tttgtcctcc
     6061 agatcgcagc aatcgcgccc ctatcttggc ccgcccacct acttatgcag gtattccccg
     6121 gggtgccatt agtggttttg tgggcaagtg gtttgaccgc agtggttagc ggggttacaa
     6181 tcagccaagt tattacaccc ttattttaca gtccaaaacc gcagggcggc gtgtgggggc
     6241 tgacgcgtgc ccccactcca caatttcaaa aaaaagagtg gccacttgtc tttgtttatg
     6301 ggccccattg gcgtggagcc ccgtttaatt ttcgggggtg ttagagacaa ccagtggagt
     6361 ccgctgctgt cggcgtccac tctctttccc cttgttacaa atagagtgta acaacatggt
     6421 tcacctgtct tggtccctgc ctgggacaca tcttaataac cccagtatca tattgcacta
     6481 ggattatgtg ttgcccatag ccataaattc gtgtgagatg gacatccagt ctttacggct
     6541 tgtccccacc ccatggattt ctattgttaa agatattcag aatgtttcat tcctacacta
     6601 gtatttattg cccaaggggt ttgtgagggt tatattggtg tcatagcaca atgccaccac
     6661 tgaacccccc gtccaaattt tattctgggg gcgtcacctg aaaccttgtt ttcgagcacc
     6721 tcacatacac cttactgttc acaactcagc agttattcta ttagctaaac gaaggagaat
     6781 gaagaagcag gcgaagattc aggagagttc actgcccgct ccttgatctt cagccactgc
     6841 ccttgtgact aaaatggttc actaccctcg tggaatcctg accccatgta aataaaaccg
     6901 tgacagctca tggggtggga gatatcgctg ttccttagga cccttttact aaccctaatt
     6961 cgatagcata tgcttcccgt tgggtaacat atgctattga attagggtta gtctggatag
     7021 tatatactac tacccgggaa gcatatgcta cccgtttagg gttaacaagg gggccttata
     7081 aacactattg ctaatgccct cttgagggtc cgcttatcgg tagctacaca ggcccctctg
     7141 attgacgttg gtgtagcctc ccgtagtctt cctgggcccc tgggaggtac atgtcggccg
     7201 caaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     7261 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     7321 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     7381 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     7441 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     7501 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     7561 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7621 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7681 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7741 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7801 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7861 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7921 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7981 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     8041 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     8101 gcctctcccc gcgcgttggc cgattcatta atg
//
