LOCUS       sequence-208153-        6887 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of vsv-CenpB 1-166-Sgo1 N61I-EGFP (no PP2A binding).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6887)
  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 6887)
  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-CenpB 1-166-Sgo1 N61I-GFP
FEATURES             Location/Qualifiers
     source          1..6887
                     /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      2827..2836
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2833..2835,2836..2838,2839..3552)
                     /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(2833..2835,2836..2838,2839..3549)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2878..2899)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3139..3158)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3486..3507
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3675..3796
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3712..3731)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3766..3785
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3803..4258)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3940..3961)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4152..4171
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4285..4389
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4391..4748
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4391..4582
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4414..4434)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4599..4734
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4783..5577
                     /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             4783..5577
                     /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(4837..4856)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5447..5466
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5765..5784)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5809..5856
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6185..6773
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6674..6693
                     /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 ggaattcggc cccaagaggc gacagctgac gttccgggag
      721 aagtcacgga tcatccagga ggtggaggag aatccggacc tgcgcaaggg cgagatcgcg
      781 cggcgcttca acatcccgcc gtccacgctg agcacgatcc tgaagaacaa gcgcgccatc
      841 ctggcgtcgg agcgcaagta cggggtggcc tccacctgcc gcaagaccaa caagctgtct
      901 ccctacgaca agctcgaggg cttgctcatc gcctggttcc agcagatccg cgccgccggc
      961 ctgccggtca agggcatcat cctcaaggag aaggcgctgc gcatagccga ggagctgggc
     1021 atggacgact tcaccgcctc caacggctgg ctggaccgct tccgccggcg ccacggcgtg
     1081 gtgtcctgca gcggcgtggc ccgcgcccgc gcgcgaaacg ctgccccccg caccccggcg
     1141 gcgcctgcca gtccggccgc ggtgccctcg gagggcggga gcactgaatt ctgcagagcc
     1201 aaggaaagat gcctgaaaaa gtcctttcaa gatagtcttg aagacataaa gaagcgaatg
     1261 aaagagaaaa ggaataaaaa cttggcagag attggcaaac gcaggtcttt tatagctgca
     1321 ccatgccaaa taatcaccaa cacttctaca ctgctgaaaa attaccaaga caacatcaaa
     1381 atgttagttt tagctttgga aaatgaaaaa tccaaagtga aagaagccca agatatcatc
     1441 ctacagctga gaaaagaatg ttactatctc acatgtcagc tatatgcatt gaaaggaaaa
     1501 cttacatcac aacaaacagt agaacctgca caaaatcaag aaatatgttc ctctggaatg
     1561 gaccccaata gtgatgacag ctccagaaat ttatttgtga aggatttacc gcaaattcct
     1621 cttgaagaaa ctgaacttcc aggacaagga gaatcatttc aaatagaaga tcagatacct
     1681 actattcctc aagacacact gggagttgat tttgattcag gtgaagctaa gtctactgat
     1741 aatgtcttac ctagaactgt atctgttcgt agcagtttaa agaaacattg taacagtata
     1801 tgtcagtttg atagcttgga tgattttgaa accagtcatt tggcagggaa gtcttttgaa
     1861 ttcgaaagag ttggattttt agacccacta gtaaacatgc acatacctga aaatgtacaa
     1921 cacaatgctt gtcaatggag caaggaccaa gttaacttat caccaaagct gattcagcca
     1981 ggaacgttta ctaaaacaaa agaagacatt ttagaatcta aatctgaaca aactaaaagt
     2041 aagcaaagag atacacaaga aagaaaaaga gaagagaaaa gaaaagctaa caggagaaaa
     2101 tcaaaacgta tgtcaaaata taaagagaat aaaagcgaaa ataaaaaaac tgttccccaa
     2161 aaaaaaatgc acaaatctgt cagttccaat gatgcttaca attttaattt ggaagagggt
     2221 gttcatctta ctcctttccg acaaaaagtg agcaatgact ctaatagaga agaaaacaac
     2281 gagtctgaag tgagcctctg tgaatcaagt ggttcaggag atgattccga tgacctctat
     2341 ttgcccactt gcaagtacat tcagaatccc acgagcaatt cagatagacc agtcaccagg
     2401 cctctagcta aaagagcact gaaatacaca gatgaaaaag agacggaggg ttctaagcca
     2461 acaaaaactc ctaccactac accacctgaa actcagcagt cacctcatct tagcctgaag
     2521 gatatcacca atgtctcctt gtatcctgtt gtgaaaatca gaagactttc tctttctcca
     2581 aaaaagaata aagcaagccc agcagtggct ctgcctaaac gtaggtgcac agccagcgtg
     2641 aactataagg agcccaccct cgcttcgaaa ctgagaagag gggacccttt tacagatttg
     2701 tgttttttga attctcctat tttcaagcag aaaaaggatt tgagacgttc taaaaaaagt
     2761 atgaaacaaa tacaaaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca
     2821 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2881 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2941 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     3001 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     3061 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3121 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3181 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3241 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     3301 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     3361 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3421 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     3481 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3541 ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     3601 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3661 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3721 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3781 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     3841 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3901 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3961 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     4021 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     4081 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     4141 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     4201 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     4261 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4321 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4381 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     4441 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4501 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4561 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4621 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4681 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4741 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4801 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4861 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4921 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     4981 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5041 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5101 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5161 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5221 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5281 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     5341 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5401 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5461 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5521 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5581 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5641 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5701 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     5761 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5821 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     5881 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     5941 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     6001 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     6061 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6121 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6181 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6241 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6301 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6361 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6421 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6481 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6541 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6601 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6661 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6721 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6781 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6841 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
