LOCUS       sequence-65984-e        7864 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 7864)
  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 7864)
  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: pTK81_GFP-4.1R
FEATURES             Location/Qualifiers
     source          1..7864
                     /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(4279..4299)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4284..4613
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4464..4599
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4628..4675
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4694..5092
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             5145..5738
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5939..5956)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6110..6698)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6190..6209)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6876..7667,7668..7736))
                     /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     7499..7518
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7737..7841)
                     /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 cgagatgcac tgcaaggttt ctttgttgga
     2281 tgacacagtt tatgaatgtg ttgtggagaa acatgctaag ggacaagatt tgcttaaacg
     2341 agtatgtgag catctcaatc ttttggaaga agactatttt ggtctagcca tttgggataa
     2401 cgcaacctct aagacatggc tggattccgc caaagaaata aaaaagcagg ttcgtggtgt
     2461 cccttggaat tttacattta atgtaaagtt ttatccacct gacccagcac agttaacaga
     2521 agacataaca agatattatt tatgtcttca gcttcggcag gacatagttg caggacgtct
     2581 gccctgttcc tttgcaacct tagcattatt aggttcttac accatccagt ctgaactggg
     2641 agactacgac ccagaactcc atggcgtgga ttatgttagt gattttaaac tggccccgaa
     2701 tcagaccaag gaacttgaag agaaggtcat ggaactgcat aagtcataca ggtccatgac
     2761 tccagctcag gctgacttgg agtttcttga gaatgccaaa aagttgtcta tgtatggagt
     2821 tgatcttcat aaagcaaagg acttggaagg agtagatatc atcctaggtg tctgctctag
     2881 tggccttctg gtttacaaag ataagctgag aattaaccgc ttcccttggc ccaaagtgct
     2941 gaagatttct tataaacgta gtagcttttt catcaagatt cggcctggag agcaagagca
     3001 gtatgaaagt accatcggat tcaaacttcc cagttaccga gcagctaaga aattatggaa
     3061 agtctgtgta gaacatcaca cgtttttcag attgacatct acagacacca ttcccaaaag
     3121 caaatttctt gcgctaggat ccaaatttcg atacagtggc cggactcaag ctcagaccag
     3181 gcaagctagt gctctaattg acaggcctgc cccacacttc gagcgtacag caagtaaacg
     3241 ggcgtcccgg agcctcgatg gagcagcagc tgtcgattcg gcagaccgaa gtcctcggcc
     3301 cacttctgca cctgccatta ctcagggtca ggttgcagaa ggtggcgtcc tagatgcctc
     3361 tgctaaaaaa acagtggtcc ctaaagcaca gaaggaaaca gtgaaggctg aagtgaaaaa
     3421 ggaagacgag ccacctgagc aagctgagcc agagcccaca gaagcatgga agaaaaagag
     3481 agaaagacta gatggtgaaa acatttatat cagacatagc aatttaatgt tggaggattt
     3541 agacaagagt caagaggaga tcaaaaaaca tcatgccagc atcagtgagc tgaaaaagaa
     3601 cttcatggag tctgtaccag aaccacggcc tagtgaatgg gataaacgct tatccactca
     3661 ctcacccttc cgaactctta acatcaatgg gcaaatcccc acaggagaag gacctcccct
     3721 ggtgaagaca caaactgtca ccatctcaga taatgccaat gctgtgaaaa gtgaaatccc
     3781 aaccaaagac gtccctattg tccacactga gaccaagacc atcacttatg aggctgccca
     3841 gactgacgac aacagtggag acttggaccc aggagtcttg ctgacagctc aaactatcac
     3901 atctgagacc ccaagcagca ccaccacaac tcaaattacc aagactgtaa aaggtgggat
     3961 ttcagagaca cgtattgaaa agagaattgt gatcacagga gatgctgata ttgaccatga
     4021 tcaggtcctt gtacaagcca tcaaggaggc aaaggagcag cacccagaca tgtcagtgac
     4081 caaggtggtc gtccaccagg agaccgagat tgctgatgag tgaccgcggg cccgggatcc
     4141 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     4201 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattcgccag
     4261 cacagtggtc gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4321 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4381 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4441 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4501 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     4561 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg
     4621 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     4681 gaggaactaa accatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc
     4741 aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct
     4801 ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg
     4861 tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat
     4921 cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt
     4981 gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac
     5041 ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct aagcacttca
     5101 tcgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc
     5161 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa
     5221 ctgagaatag agaagttcag atcaaggtca ggaacagatg gaacagctga atatgggcca
     5281 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca
     5341 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     5401 agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg
     5461 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     5521 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     5581 cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca
     5641 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     5701 ctgagtgatt gactacccgt cagcgggggt ctttcatttc cgacttgtgg tctcgctgcc
     5761 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtcttca catgcagcat
     5821 gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc atagttgcat
     5881 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttggcgctct tccgcttcct
     5941 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6001 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6061 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6121 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6181 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6241 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6301 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6361 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6421 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6481 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6541 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6601 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6661 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6721 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6781 caaaaaggat cttcacctag atccttttaa attaaaaatg aagtttgcgg ccgcaaatca
     6841 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6901 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6961 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7021 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7081 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7141 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7201 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7261 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7321 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7381 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7441 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat
     7501 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7561 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7621 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7681 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7741 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7801 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7861 ccac
//
