LOCUS       sequence-47118-e        7221 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 7221)
  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 7221)
  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: pCB163
FEATURES             Location/Qualifiers
     source          1..7221
                     /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(3636..3656)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3641..3970
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3821..3956
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3985..4032
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4051..4449
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4502..5095
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5296..5313)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5467..6055)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5547..5566)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6233..7024,7025..7093))
                     /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     6856..6875
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7094..7198)
                     /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 cgagatggcg gaccctaaat acgccgacct
     2281 tcccggcatt gccaggaatg agccagatgt ttatgaaact agcgacctac ctgaggatga
     2341 tcaagcggag ttcgatgcgg aggagctgac aagcacaagt gtggaacaca tcattgtcaa
     2401 tcctaatgct gcctatgaca agttcaagga caagagagtg gggacaaagg gacttgattt
     2461 ctcagatcgt attggaaaaa ccaagaggac aggatatgaa tctggagaat atgagatgct
     2521 tggagagggt ctgggagtga aggagacacc ccagcaaaag taccagcgcc tactgcatga
     2581 ggtccaagag ctgacaactg aagttgaaaa aatcaagacg acagtgaagg agtcagccac
     2641 agaggagaag ctgacccctg tgttgctggc taaacagctg gcagccctga agcagcagct
     2701 ggttgcttcc cacctggaga agctgctggg accagatgct gcaatcaacc ttaccgaccc
     2761 cgatggcgcc ctggctaagc gcctactact gcagctggaa gcaacaaaga acagcaaagg
     2821 gggatcaggg ggaaaaacca ctgggacccc cccagatagc agccttgtca cttatgaact
     2881 acattctcgg cctgagcagg acaagttctc tcaagctgcc aaagtcgcag aacttgaaaa
     2941 gcgcctgaca gagctggaga cagctgtacg ttgtgatcag gatgctcaga atcccctttc
     3001 tgcaggtcta cagggagcct gtctcatgga gactgtagag ctgttgcaag caaaggtgag
     3061 cgccctagac cttgcagttt tggatcaagt ggaggctcgg ctacagagtg tcctgggaaa
     3121 ggtgaacgag attgccaagc ataaagcctc tgtagaagat gcagatacac aaagcaaggt
     3181 gcaccagcta tatgaaacta tacagcgctg gagccccatt gcctccaccc tccctgagct
     3241 ggtgcagaga cttgtcacca tcaagcagct gcacgagcaa gccatgcagt ttggtcagct
     3301 cctgacacac ttggatacca cccagcagat gattgctaat tccttgaagg acaataccac
     3361 cctcttgacc caggtgcaga caaccatgcg tgaaaacctg gccacagttg aggggaactt
     3421 tgccagcatt gatgaacgga tgaagaagct gggaaagtga gaattctgca gtcgacggta
     3481 ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat accacatttg
     3541 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3601 tgaatgcaat tcgccagcac agtggtcgac cctgtggaat gtgtgtcagt tagggtgtgg
     3661 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3721 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3781 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3841 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3901 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3961 cttttgcaaa aagcttgagc acgtgttgac aattaatcat cggcatagta tatcggcata
     4021 gtataatacg acaaggtgag gaactaaacc atggccaagc ctttgtctca agaagaatcc
     4081 accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc
     4141 gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat
     4201 tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct
     4261 ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc
     4321 ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac
     4381 agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg
     4441 gagggctaag cacttcatcg ataaaataaa agattttatt tagtctccag aaaaaggggg
     4501 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
     4561 catggaaaaa tacataactg agaatagaga agttcagatc aaggtcagga acagatggaa
     4621 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
     4681 caagaacaga tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct
     4741 gccccggctc agggccaaga acagatggtc cccagatgcg gtccagccct cagcagtttc
     4801 tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat
     4861 ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc
     4921 aataaaagag cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc
     4981 gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt
     5041 ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt tcatttccga
     5101 cttgtggtct cgctgccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg
     5161 gtcttcacat gcagcatgta tcaaaattaa tttggttttt tttcttaagt atttacatta
     5221 aatggccata gttgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5281 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5341 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5401 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5461 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5521 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5581 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5641 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5701 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5761 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5821 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5881 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     5941 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6001 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6061 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6121 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6181 tttgcggccg caaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6241 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6301 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6361 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6421 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6481 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6541 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6601 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6661 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6721 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6781 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6841 cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6901 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6961 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7021 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7081 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7141 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7201 catttccccg aaaagtgcca c
//
