LOCUS       sequence-220771-        7777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of vsv-GCN4-dCEN-INCENP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7777)
  AUTHORS   van der Horst A, Vromans MJ, Bouwman K, van der Waal MS, Hadders MA,
            Lens SM
  TITLE     Inter-domain Cooperation in INCENP Promotes Aurora B Relocation from
            Centromeres to Microtubules.
  JOURNAL   Cell Rep. 2015 Jul 21;12(3):380-7. doi: 
            10.1016/j.celrep.2015.06.038. Epub 2015 Jul 9.
  PUBMED    26166576
REFERENCE   2  (bases 1 to 7777)
  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.2015.06.038"; journalName: "Cell Rep"; date: 
            "2015-07-21- 21"; volume: "12"; issue: "3"; pages: "380-7"
COMMENT     Sequence Label: pCR3-vsv-GCN4-dCEN-INCENP
FEATURES             Location/Qualifiers
     source          1..7777
                     /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"
     CDS             753..809
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     promoter        complement(3482..3500)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3483..3500)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3520..3537)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3526..3750
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3882..4470)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3962..3981)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4799..4846
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4871..4890
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5078..5872)
                     /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(5078..5872)
                     /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(5189..5208)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5799..5818
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5907..6264)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5921..6056
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6073..6264
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6221..6241
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(6324..7115,7116..7184))
                     /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     6947..6966
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7185..7289)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7316..7771
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7403..7422)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7613..7634
                     /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 aattcagaat gaaacaactt gaagacaagg ttgaagaatt gctttcgaaa aattatcact
      781 tggaaaatga ggttgccaga ttaaagaaat tagttggcga acgcggtgaa ttcagcaaag
      841 agccagagct gatgcccaaa acaccttctc agaagaaccg acggaagaag agacggattt
      901 cttatgttca ggatgaaaac agagatccca tcaggagaag gttatcccgc agaaagtctc
      961 ggagcagcca gctgagctcc cgacgcctcc gcagcaagga cagtgtagag aagctggcta
     1021 cagtggtcgg ggagaacggc tccgttctgc ggcgtgtgac ccgtgctgcg gctgcagctg
     1081 ccgcggctac catggcattg gctgcacctt cttcacccac ccctgagtct cccacgatgc
     1141 tgactaagaa gcccgaggat aaccacaccc agtgccagct ggtgcctgtg gtggagatcg
     1201 gcatcagtga gcgccagaat gctgagcagc atgtcaccca gctcatgtcc accgagcctc
     1261 tgccccgcac tctgtccccg actccagctt cagccacagc tccaacctcc cagggcatcc
     1321 cgacatcaga tgaggaatca acacctaaga agtcgaaggc caggatactg gagtccatca
     1381 cagtgagctc cctgatggct acaccccagg accccaaggg tcaaggggtc gggacggggc
     1441 ggtctgcgtc taagctcagg attgcgcagg tctcccctgg cccacgggac tcgccagcct
     1501 ttccagattc tccatggcgg gagcgggtgc tggctcccat cctgccggat aacttctcca
     1561 cgcccacggg ctctcgcacg gactctcaat cggtgcggca cagcccgatc gccccgtctt
     1621 ccccgagtcc ccgagtctta gcccagaagt actctctggt ggccaaacag gaaagtgttg
     1681 tccgcagggc gagcagaagg cttgccaaga agactgccga agagccagct gcctctggcc
     1741 gcatcatctg tcacagttac ctggagaggc tcctgaatgt tgaggtgccc cagaaagttg
     1801 gttctgagca gaaggaaccc cccgaggagg ctgagcctgt ggcggcagct gagccagagg
     1861 tccctgagaa caacggaaat aactcgtggc cccacaatga cacagagatc gccaacagca
     1921 cacccaaccc gaagcctgca gccagcagcc cggaaacacc ctctgcaggg cagcaagagg
     1981 ccaagacgga ccaagcagat ggacccagag agccaccgca gagtgccagg aggaagcgca
     2041 gctacaagca ggccgtgagt gagctggacg aggagcagca cctggaggat gaggagctgc
     2101 agccccccag gagcaagacc ccttcctcac cctgcccagc cagcaaggtg gtacggcccc
     2161 tccggacctt tctgcacaca gtgcagagga accagatgct cacgaccccg acctcagccc
     2221 cacgcagcgt catgaagtcc tttattaagc gcaacactcc cctgcgcatg gaccccaagt
     2281 gcagcttcgt cgagaaggag cggcagcgcc tggagaatct gcggcggaag gaggaggccg
     2341 agcagctgcg caggcagaag gtggaggagg acaagcggcg gcggctggag gaggtgaagc
     2401 tgaagcgtga ggaacgcctc cgcaaggtgc tgcaggcccg cgagcgggtg gagcagatga
     2461 aggaggagaa gaagaagcag attgagcaga agtttgctca gatcgacgag aagactgaga
     2521 aggccaagga ggagcggctg gcagaggaga aggccaagaa aaaggcggcg gccaagaaga
     2581 tggaggaggt ggaagcacgc aggaagcagg aagaggatgc acgtaggctc aggtggctgc
     2641 agcaggagga ggaagagcgg cggcaccaag agctgctgca gaagaagaag gaagaggagc
     2701 aggagcggct gcggaaggcg gccgaggcta agcggctggc agagcagcgg gagcaggagc
     2761 ggcgggagca ggagcggcgc gagcaggagc ggcgcgagca ggagcggcgg gagcaggagc
     2821 ggcgcgagca ggagcgacag ctggcagagc aggagcgtcg gcgggagcag gagcggctcc
     2881 aggccgagag ggagctgcag gagcgggaga aggccctgcg gctgcagaag gagcagctgc
     2941 agagggaact ggaggagaag aagaagaagg aagagcagca gcgtctggct gagcggcagc
     3001 tgcaggagga gcaagagaag aaagccaagg aggcagcagg ggccagcaag gccctgaatg
     3061 tgactgtgga cgtgcagtct ccagcttgta cctcatatca gatgactccg caagggcaca
     3121 gggcccctcc caagatcaac ccagataact acgggatgga tctgaatagc gacgactcca
     3181 ccgatgatga ggcccatccc cggaagccca tccccacctg ggcccgaggc accccgctca
     3241 gccaggctat cattcaccag tactaccacc caccgaacct tctggagctc tttggaacca
     3301 ttctcccact ggacttggag gatatcttca agaagagcaa gccccgctat cacaagcgca
     3361 ccagctctgc tgtctggaac tcaccgcccc tgcagggcgc cagggtcccc agcagcctgg
     3421 cctacagcct gaagaagcac aagctttgac tcgagtagat gactagtcta gagggcccta
     3481 ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt
     3541 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3601 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3661 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3721 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agtggcggta
     3781 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3841 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3901 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3961 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4021 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4081 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4141 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4201 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4261 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4321 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4381 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4441 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4501 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4561 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4621 gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag ccctgggcct
     4681 tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa
     4741 tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc gcgtttatga
     4801 acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat agcgcgggtt
     4861 ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac
     4921 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg
     4981 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc
     5041 gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc
     5101 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
     5161 cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat
     5221 agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca
     5281 ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca
     5341 tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca
     5401 gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt
     5461 tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat
     5521 cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg
     5581 gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg
     5641 cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat
     5701 tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc
     5761 ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc
     5821 tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg
     5881 atcctcatcc tgtctcttga tcgatctttg caaaagccta ggcctccaaa aaagcctcct
     5941 cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa
     6001 attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc
     6061 ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg
     6121 cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt
     6181 tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc
     6241 cacagctggt tctttccgcc tcaggactct tcctttttca ataaatcaat ctaaagtata
     6301 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6361 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6421 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6481 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6541 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6601 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6661 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6721 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6781 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6841 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6901 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6961 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7021 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7081 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7141 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7201 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7261 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg
     7321 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     7381 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     7441 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     7501 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     7561 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     7621 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     7681 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     7741 aattttaaca aaatattaac gcttacaatt tacgcgc
//
