LOCUS       sequence-208154-        6383 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of vsv-Sgo1 N61I-EGFP (no PP2A binding).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6383)
  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 6383)
  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: vsv-Sgo1 N61I-GFP
FEATURES             Location/Qualifiers
     source          1..6383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             649..681
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     regulatory      2323..2332
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2329..2331,2332..2334,2335..3048)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2329..2331,2332..2334,2335..3045)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2374..2395)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2635..2654)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2982..3003
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3171..3292
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3208..3227)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3262..3281
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3299..3754)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3436..3457)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3648..3667
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3781..3885
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3887..4244
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3887..4078
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3910..3930)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4095..4230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4279..5073
                     /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             4279..5073
                     /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(4333..4352)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4943..4962
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5261..5280)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5305..5352
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5681..6269
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6170..6189
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttggt accggatccg ccaccatgta caccgacatc
      661 gagatgaacc ggttgggcaa ggaattctgc agagccaagg aaagatgcct gaaaaagtcc
      721 tttcaagata gtcttgaaga cataaagaag cgaatgaaag agaaaaggaa taaaaacttg
      781 gcagagattg gcaaacgcag gtcttttata gctgcaccat gccaaataat caccaacact
      841 tctacactgc tgaaaaatta ccaagacaac atcaaaatgt tagttttagc tttggaaaat
      901 gaaaaatcca aagtgaaaga agcccaagat atcatcctac agctgagaaa agaatgttac
      961 tatctcacat gtcagctata tgcattgaaa ggaaaactta catcacaaca aacagtagaa
     1021 cctgcacaaa atcaagaaat atgttcctct ggaatggacc ccaatagtga tgacagctcc
     1081 agaaatttat ttgtgaagga tttaccgcaa attcctcttg aagaaactga acttccagga
     1141 caaggagaat catttcaaat agaagatcag atacctacta ttcctcaaga cacactggga
     1201 gttgattttg attcaggtga agctaagtct actgataatg tcttacctag aactgtatct
     1261 gttcgtagca gtttaaagaa acattgtaac agtatatgtc agtttgatag cttggatgat
     1321 tttgaaacca gtcatttggc agggaagtct tttgaattcg aaagagttgg atttttagac
     1381 ccactagtaa acatgcacat acctgaaaat gtacaacaca atgcttgtca atggagcaag
     1441 gaccaagtta acttatcacc aaagctgatt cagccaggaa cgtttactaa aacaaaagaa
     1501 gacattttag aatctaaatc tgaacaaact aaaagtaagc aaagagatac acaagaaaga
     1561 aaaagagaag agaaaagaaa agctaacagg agaaaatcaa aacgtatgtc aaaatataaa
     1621 gagaataaaa gcgaaaataa aaaaactgtt ccccaaaaaa aaatgcacaa atctgtcagt
     1681 tccaatgatg cttacaattt taatttggaa gagggtgttc atcttactcc tttccgacaa
     1741 aaagtgagca atgactctaa tagagaagaa aacaacgagt ctgaagtgag cctctgtgaa
     1801 tcaagtggtt caggagatga ttccgatgac ctctatttgc ccacttgcaa gtacattcag
     1861 aatcccacga gcaattcaga tagaccagtc accaggcctc tagctaaaag agcactgaaa
     1921 tacacagatg aaaaagagac ggagggttct aagccaacaa aaactcctac cactacacca
     1981 cctgaaactc agcagtcacc tcatcttagc ctgaaggata tcaccaatgt ctccttgtat
     2041 cctgttgtga aaatcagaag actttctctt tctccaaaaa agaataaagc aagcccagca
     2101 gtggctctgc ctaaacgtag gtgcacagcc agcgtgaact ataaggagcc caccctcgct
     2161 tcgaaactga gaagagggga cccttttaca gatttgtgtt ttttgaattc tcctattttc
     2221 aagcagaaaa aggatttgag acgttctaaa aaaagtatga aacaaataca aaagcttcga
     2281 attctgcagt cgacggtacc gcgggcccgg gatccaccgg tcgccaccat ggtgagcaag
     2341 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2401 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     2461 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2521 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     2581 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2641 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2701 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2761 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     2821 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     2881 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     2941 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     3001 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcgac
     3061 tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     3121 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     3181 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3241 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa
     3301 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3361 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3421 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     3481 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3541 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3601 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3661 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3721 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     3781 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3841 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     3901 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3961 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4021 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4081 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4141 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4201 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     4261 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4321 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4381 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4441 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     4501 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4561 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4621 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4681 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4741 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4801 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4861 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4921 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4981 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5041 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5101 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5161 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5221 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     5281 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     5341 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     5401 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     5461 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     5521 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5581 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5641 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5701 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5761 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5821 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5881 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5941 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6001 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6061 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6121 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6181 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6241 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6301 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6361 ataaccgtat taccgccatg cat
//
