LOCUS       sequence-46975-e        8029 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 8029)
  AUTHORS   Kiyomitsu T, Cheeseman IM
  TITLE     Chromosome- and spindle-pole-derived signals generate an intrinsic 
            code for spindle position and orientation.
  JOURNAL   Nat Cell Biol. 2012 Feb 12;14(3):311-7. doi: 10.1038/ncb2440.
  PUBMED    22327364
REFERENCE   2  (bases 1 to 8029)
  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"; journalName: 
            "Nat Cell Biol"; date: "2012-02-12- 12"; volume: "14"; issue: "3"; 
            pages: "311-7"
COMMENT     Sequence Label: pTK14
FEATURES             Location/Qualifiers
     source          1..8029
                     /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(4444..4464)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4449..4778
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4629..4764
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4793..4840
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4859..5257
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             5310..5903
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6104..6121)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6275..6863)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6355..6374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7041..7832,7833..7901))
                     /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     7664..7683
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7902..8006)
                     /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 cgagatgaga gaagaccatt cttttcatgt
     2281 tcgttacaga atggaagctt cttgcctaga gctggccttg gaaggggaac gtctatgtaa
     2341 atcaggagac tgccgcgctg gcgtgtcatt ctttgaagct gcagttcaag ttggaactga
     2401 agacctaaaa acacttagcg ctatttacag ccagttgggc aatgcttatt tctatttgca
     2461 tgattatgcc aaagcattag aatatcacca tcatgattta acccttgcaa ggactattgg
     2521 agaccagctg ggggaagcga aagctagtgg taatctggga aacaccttaa aagttcttgg
     2581 gaattttgac gaagccatag tttgttgtca gcgacaccta gatatttcca gagagcttaa
     2641 tgacaaggtg ggagaagcaa gagcacttta caatcttggg aatgtgtatc atgccaaagg
     2701 gaaaagtttt ggttgccctg gtccccagga tgtaggagaa tttccagaag aagtgagaga
     2761 tgctctgcag gcagccgtgg atttttatga ggaaaaccta tcattagtga ctgctttggg
     2821 tgaccgagcg gcacaaggac gtgcctttgg aaatcttgga aacacacatt acctccttgg
     2881 caacttcagg gatgcagtta tagctcatga gcagcgtctc cttattgcaa aagaatttgg
     2941 agataaagca gctgaaagaa gagcatatag caaccttgga aatgcatata tatttcttgg
     3001 tgaatttgaa actgcctcgg aatactacaa gaagacacta ctgttggccc gacagcttaa
     3061 agaccgagct gtagaagcac agtcttgtta cagtcttgga aatacatata ctttacttca
     3121 agactatgaa aaggccattg attatcatct gaagcactta gcaattgctc aagagctgaa
     3181 tgatagaatt ggtgaaggaa gagcatgttg gagcttagga aatgcataca cagcactagg
     3241 aaatcatgat caagcaatgc attttgctga aaagcacttg gaaatttcaa gagaggttgg
     3301 ggataaaagt ggtgaactaa cagcacgact taatctctca gaccttcaaa tggttcttgg
     3361 tctgagctac agcacaaata actccataat gtctgaaaat actgaaattg atagcagttt
     3421 gaatggtgta cgccccaagt tgggacgccg gcatagtatg gaaaatatgg aacttatgaa
     3481 gttaacacca gaaaaggtac agaactggaa cagtgaaatt cttgctaagc aaaaacctct
     3541 tattgccaaa ccttctgcaa agctactctt tgtcaacaga ctgaagggga aaaaatacaa
     3601 aacgaattcc tccactaaag ttctccaaga tgccagtaat tctattgacc accgaattcc
     3661 aaattctcag aggaaaatca gtgcagatac tattggagat gaagggttct ttgacttatt
     3721 aagccgattt caaagcaata ggatggatga tcagagatgt tgcttacaag aaaagaactg
     3781 ccatacagct tcaacaacaa cttcttccac tccccctaaa atgatgctaa aaacatcatc
     3841 tgttcctgtg gtatccccca acacggatga gtttttagat cttcttgcca gctcacagag
     3901 tcgccgtctg gatgaccaga gggctagttt cagtaatttg ccagggcttc gtctaacaca
     3961 aaacagccag tcggtactta gccacctgat gactaatgac aacaaagagg ctgatgaaga
     4021 tttctttgac atccttgtaa aatgtcaagg atccagatta gatgatcaaa gatgtgctcc
     4081 accacctgct accacaaagg gtccgacagt accagatgaa gactttttca gccttatttt
     4141 acggtcccag ggaaagagaa tggatgaaca gagagttctt ttacaaagag atcaaaacag
     4201 agacactgac tttgggctaa aggacttttt gcaaaataat gctttgttgg agtttaaaaa
     4261 ttcagggaaa aaatcggcag accattagcc gcgggcccgg gatccaccgg atctagataa
     4321 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     4381 cacctccccc tgaacctgaa acataaaatg aatgcaattc gccagcacag tggtcgaccc
     4441 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4501 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4561 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4621 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4681 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4741 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttgagcac gtgttgacaa
     4801 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
     4861 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
     4921 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg
     4981 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
     5041 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
     5101 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
     5161 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
     5221 tgaattgctg ccctctggtt atgtgtggga gggctaagca cttcatcgat aaaataaaag
     5281 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag
     5341 ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag
     5401 ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     5461 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca
     5521 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     5581 cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     5641 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     5701 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc
     5761 gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca
     5821 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
     5881 cccgtcagcg ggggtctttc atttccgact tgtggtctcg ctgccttggg agggtctcct
     5941 ctgagtgatt gactacccgt cagcgggggt cttcacatgc agcatgtatc aaaattaatt
     6001 tggttttttt tcttaagtat ttacattaaa tggccatagt tgcattaatg aatcggccaa
     6061 cgcgcgggga gaggcggttt gcgtattggc gctcttccgc ttcctcgctc actgactcgc
     6121 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6181 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6241 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6301 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6361 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6421 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6481 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6541 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6601 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6661 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     6721 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6781 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6841 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6901 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6961 cctagatcct tttaaattaa aaatgaagtt tgcggccgca aatcaatcta aagtatatat
     7021 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7081 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7141 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7201 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7261 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7321 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7381 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7441 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7501 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7561 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7621 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat
     7681 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7741 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7801 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7861 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7921 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7981 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccac
//
