LOCUS       sequence-65986-e        9259 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 9259)
  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 9259)
  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: pTK88_GFP-Anillin
FEATURES             Location/Qualifiers
     source          1..9259
                     /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(5674..5694)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5679..6008
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5859..5994
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6023..6070
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6089..6487
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             6540..7133
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7334..7351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7505..8093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7585..7604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8271..9062,9063..9131))
                     /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     8894..8913
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9132..9236)
                     /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 cgagatggat ccgtttacgg agaaactgct
     2281 ggagcgaacc cgtgccaggc gagagaatct tcagagaaaa atggctgaga ggcccacagc
     2341 agctccaagg tctatgactc atgctaagcg agctagacag ccactttcag aagcaagtaa
     2401 ccagcagccc ctctctggtg gtgaagagaa atcttgtaca aaaccatcgc catcaaaaaa
     2461 acgctgttct gacaacactg aagtagaagt ttctaacttg gaaaataaac aaccagttga
     2521 gtcgacatct gcaaaatctt gttctccaag tcctgtgtct cctcaggtgc agccacaagc
     2581 agcagatacc atcagtgatt ctgttgctgt cccggcatca ctgctgggca tgaggagagg
     2641 gctgaactca agattggaag caactgcagc ctcctcagtt aaaacacgta tgcaaaaact
     2701 tgcagagcaa cggcgccgtt gggataatga tgatatgaca gatgacattc ctgaaagctc
     2761 actcttctca ccaatgccat cagaggaaaa ggctgcttcc cctcccagac ctctgctttc
     2821 aaatgcctcg gcaactccag ttggcagaag gggccgtctg gccaatcttg ctgcaactat
     2881 ttgctcctgg gaagatgatg taaatcactc atttgcaaaa caaaacagtg tacaagaaca
     2941 gcctggtacc gcttgtttat ccaaattttc ctctgcaagt ggagcatctg ctaggatcaa
     3001 tagcagcagt gttaagcagg aagctacatt ctgttcccaa agggatggcg atgcctcttt
     3061 gaataaagcc ctatcctcaa gtgctgatga tgcgtctttg gttaatgcct caatttccag
     3121 ctctgtgaaa gctacttctc cagtgaaatc tactacatct atcactgatg ctaaaagttg
     3181 tgagggacaa aatcctgagc tacttccaaa aactcctatt agtcctctga aaacgggggt
     3241 atcgaaacca attgtgaagt caactttatc ccagacagtt ccatccaagg gagaattaag
     3301 tagagaaatt tgtctgcaat ctcaatctaa agacaaatct acgacaccag gaggaacagg
     3361 aattaagcct ttcctggaac gctttggaga gcgttgtcaa gaacatagca aagaaagtcc
     3421 agctcgtagc acaccccaca gaacccccat tattactcca aatacaaagg ccatccaaga
     3481 aaaattattc aagcaagaca catcttcatc tactacccat ttagcacaac agctcaagca
     3541 ggaacgtcaa aaagaactag catgtcttcg tggccgattt gacaagggca atatatggag
     3601 tgcagaaaaa ggcggaaact caaaaagcaa acaactagaa accaaacagg aaactcactg
     3661 tcagagcact cccctcaaaa aacaccaagg tgtttcaaaa actcagtcac ttccagtaac
     3721 agaaaaggtg accgaaaacc agataccagc caaaaattct agtacagaac ctaaagaagt
     3781 gatacgtgaa attgagatga gtgtggatga tgatgatatc aatagttcga aagtaattaa
     3841 tgacctcttc agtgatgtcc tagaggaagg tgaactagat atggagaaga gccaagagga
     3901 gatggatcaa gcattagcag aaagcagcga agaacaggaa gatgcactga atatctcctc
     3961 aatgtcttta cttgcaccat tggcacaaac agttggtgtg gtaagtccag agagtttagt
     4021 gtccacacct agactggaat tgaaagacac cagcagaagt gatgaaagtc caaaaccagg
     4081 aaaattccaa agaactcgtg tccctcgcgc tgaatctggt gatagccttg gttctgaaga
     4141 tcgtgatctt ctttacagca ttgatgcata tagatctcaa agattcaaag aaacagaacg
     4201 tccatcaata aagcaggtga ttgttcggaa ggaagatgtt acttcaaaac tggatgaaaa
     4261 aaataatgcc tttccttgtc aagttaatat caaacagaaa atgcaggaac tcaataacga
     4321 aataaatatg caacagacag tgatctatca agctagccag gctcttaact gctgtgttga
     4381 tgaagaacat ggaaaagggt ccctagaaga agctgaagca gaaagacttc ttctaattgc
     4441 aactgggaag agaacacttt tgattgatga attgaataaa ttgaagaacg aaggacctca
     4501 gaggaagaat aaggctagtc cccaaagtga atttatgcca tccaaaggat cagttacttt
     4561 gtcagaaatc cgcttgcctc taaaagcaga ttttgtctgc agtacggttc agaaaccaga
     4621 tgcagcaaat tactattact taattatact aaaagcagga gctgaaaata tggtagccac
     4681 accattagca agtacttcaa actctcttaa cggtgatgct ctgacattca ctactacatt
     4741 tactctgcaa gatgtatcca atgactttga aataaatatt gaagtttaca gcttggtgca
     4801 aaagaaagat ccctcaggcc ttgataagaa gaaaaaaaca tccaagtcca aggctattac
     4861 tccaaagcga ctcctcacat ctataaccac aaaaagcaac attcattctt cagtcatggc
     4921 cagtccagga ggtcttagtg ctgtgcgaac cagcaacttc gcccttgttg gatcttacac
     4981 attatcattg tcttcagtag gaaatactaa gtttgttctg gacaaggtcc cctttttatc
     5041 ttctttggaa ggtcatattt atttaaaaat aaaatgtcaa gtgaattcca gtgttgaaga
     5101 aagaggtttt ctaaccatat ttgaagatgt tagtggtttt ggtgcctggc atcgaagatg
     5161 gtgtgttctt tctggaaact gtatatctta ttggacttat ccagatgatg agaaacgcaa
     5221 gaatcccata ggaaggataa atctggctaa ttgtaccagt cgtcagatag aaccagccaa
     5281 cagagaattt tgtgcaagac gcaacacttt tgaattaatt actgtccgac cacaaagaga
     5341 agatgaccga gagactcttg tcagccaatg cagggacaca ctctgtgtta ccaagaactg
     5401 gctgtctgca gatactaaag aagagcggga tctctggatg caaaaactca atcaagttct
     5461 tgttgatatt cgcctctggc aacctgatgc ttgctacaaa cctattggaa agccttaacc
     5521 gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
     5581 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     5641 aatgcaattc gccagcacag tggtcgaccc tgtggaatgt gtgtcagtta gggtgtggaa
     5701 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5761 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5821 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5881 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5941 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     6001 tttgcaaaaa gcttgagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt
     6061 ataatacgac aaggtgagga actaaaccat ggccaagcct ttgtctcaag aagaatccac
     6121 cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt
     6181 cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt
     6241 tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg
     6301 caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg
     6361 acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag
     6421 tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga
     6481 gggctaagca cttcatcgat aaaataaaag attttattta gtctccagaa aaagggggga
     6541 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
     6601 tggaaaaata cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca
     6661 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     6721 agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     6781 cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta
     6841 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     6901 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     6961 taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg
     7021 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
     7081 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttccgact
     7141 tgtggtctcg ctgccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
     7201 cttcacatgc agcatgtatc aaaattaatt tggttttttt tcttaagtat ttacattaaa
     7261 tggccatagt tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggc
     7321 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     7381 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     7441 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     7501 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     7561 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     7621 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     7681 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     7741 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     7801 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7861 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7921 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     7981 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     8041 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     8101 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     8161 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     8221 tgcggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     8281 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     8341 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     8401 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     8461 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     8521 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     8581 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     8641 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8701 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8761 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8821 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8881 gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8941 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     9001 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     9061 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     9121 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     9181 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     9241 tttccccgaa aagtgccac
//
