LOCUS       sequence-208772-        6643 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of vsv-Sgo1 N61I (no PP2A binding).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6643)
  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 6643)
  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-Sgo1 N61I
FEATURES             Location/Qualifiers
     source          1..6643
                     /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(2348..2366)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2349..2366)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2386..2403)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2392..2616
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2748..3336)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2828..2847)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3665..3712
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3737..3756
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3944..4738)
                     /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(3944..4738)
                     /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(4055..4074)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4665..4684
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4773..5130)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4787..4922
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4939..5130
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5087..5107
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(5190..5981,5982..6050))
                     /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     5813..5832
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6051..6155)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6182..6637
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6269..6288)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6479..6500
                     /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 aattctgcag agccaaggaa agatgcctga aaaagtcctt tcaagatagt cttgaagaca
      781 taaagaagcg aatgaaagag aaaaggaata aaaacttggc agagattggc aaacgcaggt
      841 cttttatagc tgcaccatgc caaataatca ccaacacttc tacactgctg aaaaattacc
      901 aagacaacat caaaatgtta gttttagctt tggaaaatga aaaatccaaa gtgaaagaag
      961 cccaagatat catcctacag ctgagaaaag aatgttacta tctcacatgt cagctatatg
     1021 cattgaaagg aaaacttaca tcacaacaaa cagtagaacc tgcacaaaat caagaaatat
     1081 gttcctctgg aatggacccc aatagtgatg acagctccag aaatttattt gtgaaggatt
     1141 taccgcaaat tcctcttgaa gaaactgaac ttccaggaca aggagaatca tttcaaatag
     1201 aagatcagat acctactatt cctcaagaca cactgggagt tgattttgat tcaggtgaag
     1261 ctaagtctac tgataatgtc ttacctagaa ctgtatctgt tcgtagcagt ttaaagaaac
     1321 attgtaacag tatatgtcag tttgatagct tggatgattt tgaaaccagt catttggcag
     1381 ggaagtcttt tgaattcgaa agagttggat ttttagaccc actagtaaac atgcacatac
     1441 ctgaaaatgt acaacacaat gcttgtcaat ggagcaagga ccaagttaac ttatcaccaa
     1501 agctgattca gccaggaacg tttactaaaa caaaagaaga cattttagaa tctaaatctg
     1561 aacaaactaa aagtaagcaa agagatacac aagaaagaaa aagagaagag aaaagaaaag
     1621 ctaacaggag aaaatcaaaa cgtatgtcaa aatataaaga gaataaaagc gaaaataaaa
     1681 aaactgttcc ccaaaaaaaa atgcacaaat ctgtcagttc caatgatgct tacaatttta
     1741 atttggaaga gggtgttcat cttactcctt tccgacaaaa agtgagcaat gactctaata
     1801 gagaagaaaa caacgagtct gaagtgagcc tctgtgaatc aagtggttca ggagatgatt
     1861 ccgatgacct ctatttgccc acttgcaagt acattcagaa tcccacgagc aattcagata
     1921 gaccagtcac caggcctcta gctaaaagag cactgaaata cacagatgaa aaagagacgg
     1981 agggttctaa gccaacaaaa actcctacca ctacaccacc tgaaactcag cagtcacctc
     2041 atcttagcct gaaggatatc accaatgtct ccttgtatcc tgttgtgaaa atcagaagac
     2101 tttctctttc tccaaaaaag aataaagcaa gcccagcagt ggctctgcct aaacgtaggt
     2161 gcacagccag cgtgaactat aaggagccca ccctcgcttc gaaactgaga agaggggacc
     2221 cttttacaga tttgtgtttt ttgaattctc ctattttcaa gcagaaaaag gatttgagac
     2281 gttctaaaaa aagtatgaaa caaatacaaa agcttctcga gtagatgact agtctagagg
     2341 gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt
     2401 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     2461 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2521 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     2581 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagtg
     2641 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2701 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2761 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2821 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2881 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2941 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3001 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3061 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3121 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3181 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3241 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3301 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3361 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3421 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3481 atatatgagt aacctgaggc tatggcaggg cctgccgccc cgacgttggc tgcgagccct
     3541 gggccttcac ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg
     3601 ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt
     3661 ttatgaacaa acgacccaac accgtgcgtt ttattctgtc tttttattgc cgtcatagcg
     3721 cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccct agggtgggcg
     3781 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
     3841 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg
     3901 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
     3961 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     4021 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     4081 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     4141 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     4201 cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     4261 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     4321 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     4381 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     4441 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     4501 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca
     4561 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     4621 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     4681 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
     4741 gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa agcctaggcc tccaaaaaag
     4801 cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata
     4861 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
     4921 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata
     4981 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc
     5041 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac
     5101 acattccaca gctggttctt tccgcctcag gactcttcct ttttcaataa atcaatctaa
     5161 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5221 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5281 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5341 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5401 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5461 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5521 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5581 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5641 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5701 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5761 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5821 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5881 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5941 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6001 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6061 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6121 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6181 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     6241 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     6301 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     6361 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     6421 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6481 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     6541 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     6601 aacgcgaatt ttaacaaaat attaacgctt acaatttacg cgc
//
