LOCUS       sequence-208773-        7147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of vsv-CenpB 1-166-Sgo1 N61I (no PP2A binding).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7147)
  AUTHORS   Meppelink A, Kabeche L, Vromans MJ, Compton DA, Lens SM
  TITLE     Shugoshin-1 balances Aurora B kinase activity via PP2A to promote 
            chromosome bi-orientation.
  JOURNAL   Cell Rep. 2015 Apr 28;11(4):508-15. doi: 
            10.1016/j.celrep.2015.03.052. Epub 2015 Apr 16.
  PUBMED    25892238
REFERENCE   2  (bases 1 to 7147)
  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: 
            "10.1016/j.celrep.2015.03.052"; journalName: "Cell Rep"; date: 
            "2015-04-28- 28"; volume: "11"; issue: "4"; pages: "508-15"
COMMENT     Sequence Label: pCR3-vsv-CenpB 1-166-Sgo1 N61I
FEATURES             Location/Qualifiers
     source          1..7147
                     /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"
     promoter        complement(2852..2870)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2853..2870)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2890..2907)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2896..3120
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3252..3840)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3332..3351)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4169..4216
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4241..4260
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4448..5242)
                     /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(4448..5242)
                     /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(4559..4578)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5169..5188
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5277..5634)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5291..5426
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5443..5634
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5591..5611
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(5694..6485,6486..6554))
                     /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     6317..6336
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6555..6659)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6686..7141
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6773..6792)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6983..7004
                     /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 aattcggccc caagaggcga cagctgacgt tccgggagaa gtcacggatc atccaggagg
      781 tggaggagaa tccggacctg cgcaagggcg agatcgcgcg gcgcttcaac atcccgccgt
      841 ccacgctgag cacgatcctg aagaacaagc gcgccatcct ggcgtcggag cgcaagtacg
      901 gggtggcctc cacctgccgc aagaccaaca agctgtctcc ctacgacaag ctcgagggct
      961 tgctcatcgc ctggttccag cagatccgcg ccgccggcct gccggtcaag ggcatcatcc
     1021 tcaaggagaa ggcgctgcgc atagccgagg agctgggcat ggacgacttc accgcctcca
     1081 acggctggct ggaccgcttc cgccggcgcc acggcgtggt gtcctgcagc ggcgtggccc
     1141 gcgcccgcgc gcgaaacgct gccccccgca ccccggcggc gcctgccagt ccggccgcgg
     1201 tgccctcgga gggcgggagc actgaattct gcagagccaa ggaaagatgc ctgaaaaagt
     1261 cctttcaaga tagtcttgaa gacataaaga agcgaatgaa agagaaaagg aataaaaact
     1321 tggcagagat tggcaaacgc aggtctttta tagctgcacc atgccaaata atcaccaaca
     1381 cttctacact gctgaaaaat taccaagaca acatcaaaat gttagtttta gctttggaaa
     1441 atgaaaaatc caaagtgaaa gaagcccaag atatcatcct acagctgaga aaagaatgtt
     1501 actatctcac atgtcagcta tatgcattga aaggaaaact tacatcacaa caaacagtag
     1561 aacctgcaca aaatcaagaa atatgttcct ctggaatgga ccccaatagt gatgacagct
     1621 ccagaaattt atttgtgaag gatttaccgc aaattcctct tgaagaaact gaacttccag
     1681 gacaaggaga atcatttcaa atagaagatc agatacctac tattcctcaa gacacactgg
     1741 gagttgattt tgattcaggt gaagctaagt ctactgataa tgtcttacct agaactgtat
     1801 ctgttcgtag cagtttaaag aaacattgta acagtatatg tcagtttgat agcttggatg
     1861 attttgaaac cagtcatttg gcagggaagt cttttgaatt cgaaagagtt ggatttttag
     1921 acccactagt aaacatgcac atacctgaaa atgtacaaca caatgcttgt caatggagca
     1981 aggaccaagt taacttatca ccaaagctga ttcagccagg aacgtttact aaaacaaaag
     2041 aagacatttt agaatctaaa tctgaacaaa ctaaaagtaa gcaaagagat acacaagaaa
     2101 gaaaaagaga agagaaaaga aaagctaaca ggagaaaatc aaaacgtatg tcaaaatata
     2161 aagagaataa aagcgaaaat aaaaaaactg ttccccaaaa aaaaatgcac aaatctgtca
     2221 gttccaatga tgcttacaat tttaatttgg aagagggtgt tcatcttact cctttccgac
     2281 aaaaagtgag caatgactct aatagagaag aaaacaacga gtctgaagtg agcctctgtg
     2341 aatcaagtgg ttcaggagat gattccgatg acctctattt gcccacttgc aagtacattc
     2401 agaatcccac gagcaattca gatagaccag tcaccaggcc tctagctaaa agagcactga
     2461 aatacacaga tgaaaaagag acggagggtt ctaagccaac aaaaactcct accactacac
     2521 cacctgaaac tcagcagtca cctcatctta gcctgaagga tatcaccaat gtctccttgt
     2581 atcctgttgt gaaaatcaga agactttctc tttctccaaa aaagaataaa gcaagcccag
     2641 cagtggctct gcctaaacgt aggtgcacag ccagcgtgaa ctataaggag cccaccctcg
     2701 cttcgaaact gagaagaggg gaccctttta cagatttgtg ttttttgaat tctcctattt
     2761 tcaagcagaa aaaggatttg agacgttcta aaaaaagtat gaaacaaata caaaagcttc
     2821 tcgagtagat gactagtcta gagggcccta ttctatagtg tcacctaaat gctagagctc
     2881 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2941 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     3001 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     3061 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     3121 cttctgaggc ggaaagaacc agtggcggta atacggttat ccacagaatc aggggataac
     3181 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3241 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3301 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3361 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3421 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3481 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3541 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3601 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3661 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3721 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3781 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3841 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3901 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3961 tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc agggcctgcc
     4021 gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg gggtggggaa
     4081 aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc gacagagtgc
     4141 cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg cgttttattc
     4201 tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca
     4261 gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc
     4321 atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg
     4381 gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga
     4441 gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag
     4501 cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa
     4561 tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt
     4621 cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat
     4681 gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg
     4741 ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca
     4801 tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg
     4861 gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag
     4921 caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc
     4981 ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg
     5041 atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa
     5101 aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg
     5161 tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt
     5221 gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcgatctttg
     5281 caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag
     5341 gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg
     5401 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
     5461 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     5521 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
     5581 gactttccac accctaactg acacacattc cacagctggt tctttccgcc tcaggactct
     5641 tcctttttca ataaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5701 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5761 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5821 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5881 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5941 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6001 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6061 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6121 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6181 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6241 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6301 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6361 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6421 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6481 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6541 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6601 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6661 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     6721 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     6781 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     6841 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     6901 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     6961 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     7021 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     7081 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     7141 tacgcgc
//
