LOCUS       sequence-208705-        5584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of vsv-CenpB 1-166.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5584)
  AUTHORS   Liu D, Vader G, Vromans MJ, Lampson MA, Lens SM
  TITLE     Sensing chromosome bi-orientation by spatial separation of aurora B 
            kinase from kinetochore substrates.
  JOURNAL   Science. 2009 Mar 6;323(5919):1350-3. doi: 10.1126/science.1167000. 
            Epub 2009 Jan 15.
  PUBMED    19150808
REFERENCE   2  (bases 1 to 5584)
  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: 
            "10.1126/science.1167000"; journalName: "Science"; date: "2009-03-6-
            6"; volume: "323"; issue: "5919"; pages: "1350-3"
COMMENT     Sequence Label: pCR3-vsv-CenpB 1-166
FEATURES             Location/Qualifiers
     source          1..5584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     638..657
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             691..723
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     promoter        complement(1293..1311)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1294..1311)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1331..1348)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1337..1561
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(1693..2281)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1773..1792)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2610..2657
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2682..2701
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2889..3683)
                     /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(2889..3683)
                     /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(3000..3019)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3610..3629
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3718..4075)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3732..3867
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3884..4075
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4032..4052
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4135..4926,4927..4995))
                     /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     4758..4777
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4996..5100)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5127..5582
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5214..5233)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5424..5445
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      601 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      661 cccaagcttg gtaccggatc cgccaccatg tacaccgaca tcgagatgaa ccggttgggc
      721 aaggaattcg gccccaagag gcgacagctg acgttccggg agaagtcacg gatcatccag
      781 gaggtggagg agaatccgga cctgcgcaag ggcgagatcg cgcggcgctt caacatcccg
      841 ccgtccacgc tgagcacgat cctgaagaac aagcgcgcca tcctggcgtc ggagcgcaag
      901 tacggggtgg cctccacctg ccgcaagacc aacaagctgt ctccctacga caagctcgag
      961 ggcttgctca tcgcctggtt ccagcagatc cgcgccgccg gcctgccggt caagggcatc
     1021 atcctcaagg agaaggcgct gcgcatagcc gaggagctgg gcatggacga cttcaccgcc
     1081 tccaacggct ggctggaccg cttccgccgg cgccacggcg tggtgtcctg cagcggcgtg
     1141 gcccgcgccc gcgcgcgaaa cgctgccccc cgcaccccgg cggcgcctgc cagtccggcc
     1201 gcggtgccct cggagggcgg gagcactgaa ttctgcagat atccatcaca ctggcggccg
     1261 ctcgagtaga tgactagtct agagggccct attctatagt gtcacctaaa tgctagagct
     1321 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1381 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1441 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     1501 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     1561 gcttctgagg cggaaagaac cagtggcggt aatacggtta tccacagaat caggggataa
     1621 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1681 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1741 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1801 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1861 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1921 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1981 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2041 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2101 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2161 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2221 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2281 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2341 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2401 atgaagtttt aaatcaatct aaagtatata tgagtaacct gaggctatgg cagggcctgc
     2461 cgccccgacg ttggctgcga gccctgggcc ttcacccgaa cttggggggt ggggtgggga
     2521 aaaggaagaa acgcgggcgt attggcccca atggggtctc ggtggggtat cgacagagtg
     2581 ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt
     2641 ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct tccgtgtttc
     2701 agttagcctc cccctagggt gggcgaagaa ctccagcatg agatccccgc gctggaggat
     2761 catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt
     2821 ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag
     2881 agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga
     2941 gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca
     3001 atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag
     3061 tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca
     3121 tgggtcacga cgagatcctc gccgtcgggc atgctcgcct tgagcctggc gaacagttcg
     3181 gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc
     3241 atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc
     3301 ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga
     3361 gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt
     3421 cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac
     3481 gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc ggtcttgaca
     3541 aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt
     3601 gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg
     3661 tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcgatcttt
     3721 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     3781 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     3841 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     3901 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     3961 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     4021 ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc ctcaggactc
     4081 ttcctttttc aataaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4141 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4201 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4261 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4321 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4381 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4441 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4501 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4561 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4621 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4681 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4741 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4801 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4861 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4921 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4981 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5041 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5101 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     5161 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5221 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5281 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5341 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5401 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5461 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     5521 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     5581 ttac
//
