LOCUS       sequence-100528-        6301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Allows the integration of Myc ArB in the genome and Tet-inducible 
            expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6301)
  AUTHORS   Girdler F, Gascoigne KE, Eyers PA, Hartmuth S, Crafter C, Foote KM, 
            Keen NJ, Taylor SS
  TITLE     Validating Aurora B as an anti-cancer drug target.
  JOURNAL   J Cell Sci. 2006 Sep 1;119(Pt 17):3664-75. Epub 2006 Aug 15.
  PUBMED    16912073
REFERENCE   2  (bases 1 to 6301)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Sci."; date: "2006-09-1"; volume: "119(Pt 17)"; pages: "3664-75. 
            Epub 2006 Aug 15"
COMMENT     Sequence Label: pcDNA5 FRT TO Myc ArB WT RNAiR
FEATURES             Location/Qualifiers
     source          1..6301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1154..1183
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1916..1927
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2253..2270)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2259..2483
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2616..2635)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2767..2814
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             2822..3844
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             2822..3844
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    3974..4095
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4011..4030)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4065..4084
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4144..4160)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4144..4160)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4157..4179)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4168..4184
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4192..4198,4199..4216,4217..4222))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4237..4258
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4375..4392)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4546..5134)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4626..4645)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5305..6096,6097..6165))
                     /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     5928..5947
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6166..6270)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccc cggggaattc ggcagatccc gaggtccgac
     1021 agcgcccggc ccagatcccc acgcctgcca ggagcaagcc gagagccagc cggccggcgc
     1081 actccgactc cgagcagtct ctgtccttcg acccgagccc cgcgcccttt ccgggacccc
     1141 tgccccgcgg atggagcaga agctcatctc agaagaagac ctcgaggccc agaaggagaa
     1201 ctcctacccc tggccctacg gccgacagac ggctccatct ggcctgagca ccctgcccca
     1261 gcgagtcctc cggaaagagc ctgtcacccc atctgcactt gtcctcatga gccgctccaa
     1321 tgtccagccc acagctgccc ctggccagaa ggtgatggag aatagcagtg ggacacccga
     1381 catcttaacg cggcacttca caattgatga ctttgagatt gggcgtcctc tgggcaaagg
     1441 caagtttgga aacgtgtact tggctcggga gaagaaaagc catttcatcg tggcgctcag
     1501 ggtcctcttc aagtcccaga tagagaagga gggcgtggag catcagctgc gcagagagat
     1561 cgaaatccag gcccacctgc accatcccaa catcctgcgt ctctacaact atttttatga
     1621 ccggaggagg atctacttga ttctagagta tgccccccgc ggggagctct acaaggagct
     1681 gcagaagagc tgcacatttg acgagcagcg aacagccacg atcatggagg agttggcaga
     1741 tgctctaatg tactgccatg ggaagaaggt gattcacaga gacataaagc cagaaaatct
     1801 gctcttaggg ctcaagggag agctgaagat tgctgacttc ggctggtctg tgcatgcgcc
     1861 ctccctgagg aggaagacaa tgtgtggcac cctggactac ctgcccccag agatgattga
     1921 ggggcgcatg cacaatgaga aggtggatct gtggtgcatt ggagtgcttt gctatgagct
     1981 gctggtgggg aacccaccct ttgagagtgc atcacacaac gagacctatc gccgcatcgt
     2041 caaggtggac ctaaagttcc ccgcttctgt gcccacggga gcccaggacc tcatctccaa
     2101 actgctcagg cataacccct cggaacggct gcccctggcc caggtctcag cccacccttg
     2161 ggtccgggcc aactctcgga gggtgctgcc tccctctgcc cttcaatctg tcgcctgagg
     2221 cggccgcggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     2281 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2341 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2401 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     2461 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     2521 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2581 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2641 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2701 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2761 tacctagaag ttcctattcc gaagttccta ttctctagaa agtataggaa cttccttggc
     2821 caaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     2881 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     2941 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     3001 ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg
     3061 ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     3121 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc
     3181 gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     3241 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     3301 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     3361 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     3421 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     3481 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     3541 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg
     3601 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     3661 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     3721 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg
     3781 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     3841 atagcacgta ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     3901 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     3961 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4021 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4081 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     4141 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4201 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4261 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4321 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4381 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4441 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4501 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4561 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4621 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4681 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4741 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4801 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4861 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4921 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4981 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5041 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5101 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5161 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5221 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5281 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5341 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5401 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5461 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5521 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5581 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5641 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5701 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5761 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5821 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5881 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5941 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6001 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6061 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6121 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6181 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6241 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6301 c
//
