LOCUS       sequence-65985-e        9013 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 9013)
  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 9013)
  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: pTK82_GFP-4.1G
FEATURES             Location/Qualifiers
     source          1..9013
                     /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(5428..5448)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5433..5762
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5613..5748
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5777..5824
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5843..6241
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             6294..6887
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7088..7105)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7259..7847)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7339..7358)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8025..8816,8817..8885))
                     /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     8648..8667
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8886..8990)
                     /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 cgagatgact actgaagtag gctctgtgtc
     2281 tgaagtgaag aaggactcta gccagttagg aacagatgca accaaggaaa aacctaaaga
     2341 agtagcagaa aatcagcaga atcagtcttc cgatccagag gaggaaaaag gttcccagcc
     2401 acctcctgca gctgaaagcc aaagtagtct acgccgccag aagagagaga aggaaacatc
     2461 ggagagcagg ggtatttctc ggttcatacc gccatggctt aagaagcaaa agtcatatac
     2521 cttagtagtg gccaaagatg gaggagataa aaaagagcct acccaagctg ttgttgaaga
     2581 acaggtctta gataaagagg aaccccttcc agaagaacag agacaggcta agggtgatgc
     2641 tgaagaaatg gctcagaaga aacaagagat taaagttgaa gtcaaggaag aaaaaccctc
     2701 agtgagcaag gaagaaaaac cctcagtgag caaagtggag atgcagccta ctgaattagt
     2761 aagtaaggag agagaagaga aggtaaaaga aacacaggaa gacaaattag aaggaggagc
     2821 agcaaaaagg gagaccaagg aagtgcagac caatgagctg aaagcagaga aggcatctca
     2881 aaaagtcacc aagaagacca aaactgtcca gtgtaaagtg accctcttag atggcaccga
     2941 atacagctgt gacctggaga aacatgccaa gggacaagtg ttatttgaca aagtgtgtga
     3001 acacctcaat ctcttggaga aagactactt tggacttttg tttcaggaaa gccctgagca
     3061 gaaaaactgg ttagatcctg ctaaagaaat aaagagacaa ctgagaaacc ttccatggct
     3121 attcactttt aatgtgaagt tttatcctcc tgatccttct caattgactg aagatatcac
     3181 cagatacttc ttgtgccttc agctccggca ggacattgcc tctggccgcc tgccctgctc
     3241 ttttgtgact catgctctcc tgggatccta caccctgcag gctgaacttg gtgactatga
     3301 cccagaagaa catggcagca tcgacctcag tgaattccag tttgccccta ctcagactaa
     3361 ggagctggaa gagaaggtgg cagagctgca caaaacccac aggggcttat cgccagcaca
     3421 agctgattcc cagttcttag aaaatgcaaa gaggctttcc atgtatggtg ttgacctaca
     3481 tcatgccaag gactcagaag gtgtggacat caagctgggc gtgtgtgcta atggacttct
     3541 catttacaaa gacagactgc gaatcaatcg ttttgcttgg ccgaaaatct taaaaatttc
     3601 ctataaacgc agtaacttct acattaaagt cagaccggca gagctggaac agtttgagag
     3661 taccattgga ttcaaactgc caaaccaccg ggcagcgaaa agactatgga aagtgtgcgt
     3721 ggagcatcat actttctaca ggcttgtttc tccagagcag ccaccaaaag ccaagttcct
     3781 gaccttgggg tccaaatttc gctatagtgg ccgcacccaa gcacagaccc gccaggccag
     3841 caccctcata gataggccag caccacactt tgagcgcact tctagtaaac gggtctccag
     3901 gagtctagat ggagctccga ttggtgtcat ggaccaaagt cttatgaagg attttcctgg
     3961 cgctgctggg gagatttcag cctatggacc tggacttgtc agcattgccg tggtacaaga
     4021 tggggacggc aggagggaag tgagaagccc aactaaagcc ccacatttgc agctcattga
     4081 aggaaagaaa aattccttga gagtagaagg ggataatatt tatgtcagac atagcaattt
     4141 aatgttggag gaactggata aggcccagga ggacatactg aaacatcagg ctagcattag
     4201 tgaactcaag cgcaatttta tggaatccac acctgagccg cgccctaatg aatgggaaaa
     4261 acgacgtatc acacctctgt ccctacagac acaagggagt tcacatgaga ctctgaatat
     4321 agtggaggag aagaagcggg cagaggttgg gaaagacgaa agagtaatca cagaagaaat
     4381 gaatggtaaa gagatatcac ctgggagtgg tcctggggag attcgtaagg tggagcctgt
     4441 gacacaaaaa gactccacct ccctgtcttc tgagagcagc agcagcagca gtgagagtga
     4501 ggaggaagac gtgggagagt accgtcccca ccaccgagtg accgagggca ccatcaggga
     4561 ggaacaggag tatgaagaag aggtggagga agaaccccgc ccggcagcca aggtagtaga
     4621 gagggaggaa gcagtgcccg aagccagccc agtcacacaa gcaggtgcca gtgtaatcac
     4681 agtagaaaca gtgatccagg aaaatgtagg tgcccaaaag atacccggag agaagagtgt
     4741 acacgaaggc gctcttaagc aagacatggg agaagaagca gaggaagagc cacagaaagt
     4801 taacggagag gtgtcccatg ttgacattga tgttttgcca caaattattt gttgttcaga
     4861 gccaccagtg gtaaaaacag agatggtaac aatttctgat gcctcacaaa ggacagaaat
     4921 ctccaccaag gaagtcccca ttgtccaaac tgagaccaaa accatcacat atgagtctcc
     4981 acagattgat ggcggggctg gtggtgattc gggcacgtta ctgaccgcac aaaccatcac
     5041 atctgagtcc gtgtcaacaa cgacaaccac acacatcacc aagactgtaa aaggtggaat
     5101 ttctgaaaca agaattgaga aacgcattgt gatcacagga gatggagata ttgatcatga
     5161 ccaggcactg gctcaggcga tcagggaagc cagagagcag caccctgaca tgtcggtcac
     5221 aagagtggtg gtacacaaag aaacagagtt ggctgaggaa ggggaagatt aaccgcgggc
     5281 ccgggatcca ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt
     5341 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     5401 attcgccagc acagtggtcg accctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     5461 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     5521 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5581 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     5641 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     5701 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     5761 aaaagcttga gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     5821 cgacaaggtg aggaactaaa ccatggccaa gcctttgtct caagaagaat ccaccctcat
     5881 tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag
     5941 cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg
     6001 gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct
     6061 gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg
     6121 ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg
     6181 acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcta
     6241 agcacttcat cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa
     6301 gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa
     6361 aatacataac tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa
     6421 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     6481 gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     6541 tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac
     6601 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
     6661 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
     6721 agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc
     6781 gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga
     6841 gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttcc gacttgtggt
     6901 ctcgctgcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtcttcac
     6961 atgcagcatg tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca
     7021 tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tggcgctctt
     7081 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     7141 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     7201 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7261 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7321 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     7381 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     7441 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     7501 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     7561 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     7621 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7681 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7741 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     7801 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7861 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7921 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agtttgcggc
     7981 cgcaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     8041 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     8101 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     8161 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     8221 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     8281 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     8341 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     8401 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     8461 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     8521 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     8581 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     8641 acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     8701 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     8761 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     8821 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     8881 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     8941 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     9001 cgaaaagtgc cac
//
