LOCUS       sequence-65828-e        8955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8955)
  AUTHORS   Kiyomitsu T, Cheeseman IM
  TITLE     Cortical dynein and asymmetric membrane elongation coordinately 
            position the spindle in anaphase.
  JOURNAL   Cell. 2013 Jul 18;154(2):391-402. doi: 10.1016/j.cell.2013.06.010.
  PUBMED    23870127
REFERENCE   2  (bases 1 to 8955)
  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.cell.2013.06"; journalName: "Cell"; date: "2013-07-18- 
            18"; volume: "154"; issue: "2"; pages: "391-402"
COMMENT     Sequence Label: pTK64_GFP-NuMA-Bonsai
FEATURES             Location/Qualifiers
     source          1..8955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             934..1350
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1318..1334
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1385..1394
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1391..1393,1394..1396,1397..2107)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1391..1393,1394..1396,1397..2107)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1436..1457)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1697..1716)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2044..2065
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2153..2173
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2186..2230
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(5370..5390)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5375..5704
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5555..5690
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5719..5766
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5785..6183
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             6236..6829
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7030..7047)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7201..7789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7281..7300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7967..8758,8759..8827))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8590..8609
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8828..8932)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt
      901 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt
      961 ttgaccttag atcactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc
     1021 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg
     1081 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca
     1141 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct
     1201 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttctt
     1261 ccatccgcgc cgtctctccc ccttgaacct cctctttcga ccccgcctca atcctccctt
     1321 tatccagccc tcactccttc tctaggcgcc ggccggatcc cagtgtggtg gtacgctacc
     1381 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1441 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     1501 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1561 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     1621 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     1681 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     1741 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1801 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     1861 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     1921 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1981 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     2041 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2101 gtacaagtcc ggaggggatc ggtggagctc caccggtggc ggccgctcta gagagaatct
     2161 ttattttcag ggcgccgcca aattcaaaga aaccgctgct gctaaattcg aacgccagca
     2221 catggacagc ggaggtggag gtactagtct cgacatgaca ctccacgcca cccggggggc
     2281 tgcactcctc tcttgggtga acagtctaca cgtggctgac cctgtggagg ctgtgctgca
     2341 gctccaggac tgcagcatct tcatcaagat cattgacaga atccatggca ctgaagaggg
     2401 acagcaaatc ttgaagcagc cggtgtcaga gagactggac tttgtgtgca gttttctgca
     2461 gaaaaatcga aaacatccct cttccccaga atgcctggta tctgcacaga aggtgctaga
     2521 gggatcagag ctggaactgg cgaagatgac catgctgctc ttataccact ctaccatgag
     2581 ctccaaaagt cccagggact gggaacagtt tgaatataaa attcaggctg agttggctgt
     2641 cattcttaaa tttgtgctgg accatgagga cgggctaaac cttaatgagg acctagagaa
     2701 cttcctacag aaagctcctg tgccttctac ctgttctagc acattccctg aagagctctc
     2761 cccacctagc caccaggcca agagggagat tcgcttccta gagctacaga aggttgcctc
     2821 ctcttccagt gggaacaact ttctctcagg ttctccagct tctcccatgg gtgatatcct
     2881 gcagacccca cagttccaga tgagacggct gaagaagcag cttgctgatg agagaagtaa
     2941 tagggatgag ctggagctgg agctagctga gaaccgcaag ctcctcaccg agaaggatgc
     3001 acagatagcc atgatgcagc agcgcattga ccgcctagcc ctgctgaatg agaagcaggc
     3061 ggccagccca ctggagccca aggagcttga ggagctgcgt gacaagaatg agagccttac
     3121 catgcggctg catgaaaccc tgaagcagtg ccaggacctg aagacagaga agagccagat
     3181 ggatcgcaaa atcaaccagc tttcggagga gaatggagac ctttccttta agctgcggga
     3241 gtttgccagt catctgcagc agctacagga tgccctcaat gagctgacgg aggagcacag
     3301 caaggccact caggagtggc tagagaagca ggcccagctg gagaaggagc tcagcgcagc
     3361 cctgcaggac aagaaatgcc ttgaagagaa gaacgaaatc cttcagggaa aactttcaca
     3421 gctggaagaa cacttgtccc agctgcagga taacccaccc caggagaagg gcgaggtgct
     3481 gggtgatgtc ttgcaggtgg aagaactgag taagaaactg gctgactctg accaagccag
     3541 caaggtgcag cagcagaagc tgaaggctgt ccaggctcag ggaggcgaga gccagcagga
     3601 ggcccagcgc ctccaggccc agctgaatga actgcaagcc cagttgagcc agaaggagca
     3661 ggcagctgag cactataagc tgcagatgga gaaagccaaa acacattatg atgccaagaa
     3721 gcagcagaac caagagctgc aggagcagct gcggagcctg gagcagctgc agaaggaaaa
     3781 caaagagctg cgagctgaag ctgaacggct gggccatgag ctacagcagg ctgggctgaa
     3841 gaccaaggag gctgaacaga cctgccgcca ccttactgcc caggtgcgca gcctggaggc
     3901 acaggttgcc catgcagacc agcagcttcg agacctgggc aaattccagg tggcaactga
     3961 tgctttaaag agccgtgagc cccaggctaa gccccagctg gacttgagta ttgacagcct
     4021 ggatctgagc tgcgaggagg ggaccccact cagtatcacc agcaagctgc ctcgtaccca
     4081 gccagacggc accagcgtcc ctggagaacc agcctcacct atctcccagc gcctgccccc
     4141 caaggtagaa tccctggaga gtctctactt cactcccatc cctgctcgga gtcaggcccc
     4201 cctggagagc agcctggact ccctgggaga cgtcttcctg gactcgggtc gtaagacccg
     4261 ctccgctcgt cggcgcacca cgcagatcat caacatcacc atgaccaaga agctagatgt
     4321 ggaagagcca gacagcgcca actcatcgtt ctacagcacg cggtctgctc ctgcttccca
     4381 ggctagcctg cgagccacct cctctactca gtctctagct cgcctgggtt ctcccgatta
     4441 tggcaactca gccctgctca gcttgcctgg ctaccgcccc accactcgca gttctgctcg
     4501 tcgttcccag gccggggtgt ccagtggggc ccctccagga aggaacagct tctacatggg
     4561 cacttgccag gatgagcctg agcagctgga tgactggaac cgcattgcag agctgcagca
     4621 gcgcaatcga gtgtgccccc cacatctgaa gacctgctat cccctggagt ccaggccttc
     4681 cctgagcctg ggcaccatca cagatgagga gatgaaaact ggagaccccc aagagaccct
     4741 gcgccgagcc agcatgcagc caatccagat agccgagggc actggcatca ccacccggca
     4801 gcagcgcaaa cgggtctccc tagagcccca ccagggccct ggaactcctg agtctaagaa
     4861 ggccaccagc tgtttcccac gccccatgac tccccgagac cgacatgaag ggcgcaaaca
     4921 gagcactact gaggcccaga agaaagcagc tccagcttct actaaacagg ctgaccggcg
     4981 ccagtcgatg gccttcagca tcctcaacac acccaagaag ctagggaaca gccttctgcg
     5041 gcggggagcc tcaaagaagg ccctgtccaa ggcttccccc aacactcgca gtggaacccg
     5101 ccgttctccg cgcattgcca ccaccacagc cagcgccgcc actgctgccg ccattggtgc
     5161 cacccctcga gccaagggca aggcaaagca ctaagaattc tgcagtcgac ggtaccgcgg
     5221 gcccgggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg
     5281 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     5341 caattcgcca gcacagtggt cgaccctgtg gaatgtgtgt cagttagggt gtggaaagtc
     5401 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     5461 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5521 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5581 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5641 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5701 caaaaagctt gagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     5761 tacgacaagg tgaggaacta aaccatggcc aagcctttgt ctcaagaaga atccaccctc
     5821 attgaaagag caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc
     5881 agcgcagctc tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact
     5941 gggggacctt gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac
     6001 ctgacttgta tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg
     6061 tgccgacagg tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat
     6121 ggacagccga cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc
     6181 taagcacttc atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     6241 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     6301 aaaatacata actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg
     6361 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     6421 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     6481 gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga
     6541 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     6601 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     6661 agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac
     6721 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     6781 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt ccgacttgtg
     6841 gtctcgctgc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttc
     6901 acatgcagca tgtatcaaaa ttaatttggt tttttttctt aagtatttac attaaatggc
     6961 catagttgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attggcgctc
     7021 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     7081 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     7141 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7201 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7261 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7321 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7381 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7441 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7501 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7561 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7621 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7681 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7741 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7801 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7861 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagtttgcg
     7921 gccgcaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7981 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     8041 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     8101 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     8161 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8221 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8281 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8341 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8401 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8461 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8521 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8581 caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8641 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8701 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8761 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8821 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8881 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8941 cccgaaaagt gccac
//
