LOCUS       sequence-208770-        6137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of vsv-AuroraB Analog sensitive (L154A/H250Y).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6137)
  AUTHORS   Hengeveld RC, Hertz NT, Vromans MJ, Zhang C, Burlingame AL, Shokat 
            KM, Lens SM
  TITLE     Development of a chemical genetic approach for human aurora B kinase
            identifies novel substrates of the chromosomal passenger complex.
  JOURNAL   Mol Cell Proteomics. 2012 May;11(5):47-59. doi: 
            10.1074/mcp.M111.013912. Epub 2012 Jan 20.
  PUBMED    22267324
REFERENCE   2  (bases 1 to 6137)
  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"; doi: 
            "10.1074/mcp.M111.013912"; journalName: "Mol Cell Proteomics"; date:
            "2012-05"; volume: "11"; issue: "5"; pages: "47-59"
COMMENT     Sequence Label: pCR3-vsv-AuroraB L154A/H250Y
FEATURES             Location/Qualifiers
     source          1..6137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     540..560
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     634..653
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        634..652
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             687..719
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     regulatory      726..735
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1464..1475
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(1842..1860)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1843..1860)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1880..1897)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1886..2110
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2242..2830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2322..2341)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3159..3206
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3231..3250
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3438..4232)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(3438..4232)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3549..3568)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4159..4178
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4267..4624)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4281..4416
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4433..4624
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4581..4601
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4684..5475,5476..5544))
                     /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     5307..5326
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5545..5649)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5676..6131
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5763..5782)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5973..5994
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      421 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      481 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      541 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
      601 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
      661 agcttggtac cggatccgcc accatgtaca ccgacatcga gatgaaccgg ttgggcaagg
      721 aattcgccac catggcccag aaggagaact cctacccctg gccctacggc cgacagacgg
      781 ctccatctgg cctgagcacc ctgccccagc gagtcctccg gaaagagcct gtcaccccat
      841 ctgcacttgt cctcatgagc cgctccaatg tccagcccac agctgcccct ggccagaagg
      901 tgatggagaa tagcagtggg acacccgaca tcttaacgcg gcacttcaca attgatgact
      961 ttgagattgg gcgtcctctg ggcaaaggca agtttggaaa cgtgtacttg gctcgggaga
     1021 agaaaagcca tttcatcgtg gcgctcaagg tcctcttcaa gtcccagata gagaaggagg
     1081 gcgtggagca tcagctgcgc agagagatcg aaatccaggc ccacctgcac catcccaaca
     1141 tcctgcgtct ctacaactat ttttatgacc ggaggaggat ctacttgatt gcagagtatg
     1201 ccccccgcgg ggagctctac aaggagctgc agaagagctg cacatttgac gagcagcgaa
     1261 cagccacgat catggaggag ttggcagatg ctctaatgta ctgccatggg aagaaggtga
     1321 ttcacagaga cataaagcca gaaaatctgc tcttagggct caagggagag ctgaagattg
     1381 ctgacttcgg ctggtctgtg catgcgccct ccctgaggag gaagacaatg tgtggcaccc
     1441 tggactacct gcccccagag atgattgagg ggcgcatgta caatgagaag gtggatctgt
     1501 ggtgcattgg agtgctttgc tatgagctgc tggtggggaa cccacccttt gagagtgcat
     1561 cacacaacga gacctatcgc cgcatcgtca aggtggacct aaagttcccc gcttctgtgc
     1621 ccacgggagc ccaggacctc atctccaaac tgctcaggca taacccctcg gaacggctgc
     1681 ccctggccca ggtctcagcc cacccttggg tccgggccaa ctctcggagg gtgctgcctc
     1741 cctctgccct tcaatctgtc gcctgacccg ggaatcgaat tctgcagata tccatcacac
     1801 tggcggccgc tcgagtagat gactagtcta gagggcccta ttctatagtg tcacctaaat
     1861 gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     1921 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     1981 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2041 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2101 ggctctatgg cttctgaggc ggaaagaacc agtggcggta atacggttat ccacagaatc
     2161 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2221 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2281 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2341 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2401 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2461 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2521 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2581 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2641 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2701 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2761 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2821 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2881 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2941 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc
     3001 agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg
     3061 gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc
     3121 gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg
     3181 cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt
     3241 ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg
     3301 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa
     3361 ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc
     3421 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     3481 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     3541 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
     3601 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     3661 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg
     3721 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     3781 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     3841 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     3901 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     3961 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     4021 gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg
     4081 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     4141 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     4201 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
     4261 tcgatctttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     4321 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     4381 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     4441 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     4501 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     4561 ggagcctggg gactttccac accctaactg acacacattc cacagctggt tctttccgcc
     4621 tcaggactct tcctttttca ataaatcaat ctaaagtata tatgagtaaa cttggtctga
     4681 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4741 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4801 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4861 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4921 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4981 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5041 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5101 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5161 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5221 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5281 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5341 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5401 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5461 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5521 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5581 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5641 ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag
     5701 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5761 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5821 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5881 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5941 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6001 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     6061 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     6121 gcttacaatt tacgcgc
//
