LOCUS       sequence-47120-e        7902 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 7902)
  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 7902)
  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: pCB168
FEATURES             Location/Qualifiers
     source          1..7902
                     /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(4317..4337)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4322..4651
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4502..4637
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4666..4713
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4732..5130
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             5183..5776
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5977..5994)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6148..6736)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6228..6247)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6914..7705,7706..7774))
                     /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     7537..7556
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7775..7879)
                     /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 cgagatgtct gacaaaagtg acttaaaagc
     2281 tgagctagag cgcaaaaagc agcgcttagc acagataaga gaagagaaga aacggaagga
     2341 agaggagagg aaaaagaaag aggctgatat gcagcagaag aaagaacccg ttcaggacga
     2401 ctctgatctg gatcgcaaac gacgagagac agaggctttg ctgcaaagca ttggtatctc
     2461 accggagccg cctctagtcc caacccctat gtctccctcc tcgaaatcag tgagcactcc
     2521 cagtgaagct ggaagccaag actcaggcga tctggggcca ttaacaagga ccctgcagtg
     2581 ggacacagac ccctcagtgc tccagctgca gtcagactca gaacttggaa gaagactgca
     2641 taaactgggc gtgtcaaagg tcacccaagt ggatttcctg ccaagggaag tagtgtccta
     2701 ctcaaaggag acccagactc ctcttgccac gcatcagtct gaagaggatg aggaagatga
     2761 ggaaatggtg gaatctaaag ttggccagga ctcagaactg gaaaatcagg acaaaaaaca
     2821 ggaagtgaag gaagcccctc caagagagtt gacagaggaa gaaaaacagc agatcattca
     2881 ttcagaggaa tttctcatct tttttgaccg gacaatacgg gtaattgaaa gagccctggc
     2941 tgaagattcc gacatctttt ttgactacag cggccgagag ttagaggaaa aagatgggga
     3001 tgttcaggct ggagccaatc tttctttcaa tcgtcagttc tatgatgaac attgttccaa
     3061 gcatcgagtg gtcacttgta tggactggtc cctccagtac cctgagctga tggtggcttc
     3121 ttacaacaac aatgaagatg ctccccatga accagatgga gtggccttgg tttggaacat
     3181 gaagtttaag aaaaccacac cagaatacgt cttccactgt cagtcctctg tgatgtcggt
     3241 ctgcttcgcc cgtttccatc ctaacttggt ggttggtggg acttactcgg gccagattgt
     3301 cctctgggac aatcgcagtc atcgaaggac tccagtgcag cggacaccct tatcagctgc
     3361 tgcacacacg catcccgtgt actgtgtaaa tgttgttggg acccagaatg ctcataacct
     3421 catcactgtc tccactgatg gcaaaatgtg ttcctggagc ctggacatgc tctcaactcc
     3481 acaggagagc atggagctgg tgtacaataa gtccaagcct gtcgctgtta ccggaatggc
     3541 tttcccaacg ggagacgtca ataacttcgt ggttggcagt gaggaaggta cagtctacac
     3601 ggcttgtcgt catggaagca aagcaggtat tggtgaggtc tttgaaggtc accaagggcc
     3661 agtgacagga attaactgcc acatggcagt gggaccaatc gacttttctc acctgtttgt
     3721 cacatcatca tttgactgga ctgtcaaact gtggaccacc aagcacaaca agccgctcta
     3781 ctcctttgaa gacaatgcag actatgtgta cgatgtcatg tggtcccccg tgcatcctgc
     3841 gctttttgcc tgcgtggacg ggatggggcg cttggacctc tggaacctca acaatgacac
     3901 cgaggttcca acagcaagtg tggccattga gggggcatcc gccctaaacc gtgttcgttg
     3961 ggcccaagct ggcaaagaag ttgctgttgg ggactcggaa ggccgtattt gggtctatga
     4021 cgttggagag cttgcagttc cccacaatga tgaatggacc cgatttgcca ggacccttgt
     4081 ggaaattcgt gctaacagag ctgatagcga ggaggaaggc actgttgagt tatctgccta
     4141 ggaattctgc agtcgacggt accgcgggcc cgggatccac cggatctaga taactgatca
     4201 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     4261 ccctgaacct gaaacataaa atgaatgcaa ttcgccagca cagtggtcga ccctgtggaa
     4321 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4381 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4441 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4501 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4561 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4621 aggctttttt ggaggcctag gcttttgcaa aaagcttgag cacgtgttga caattaatca
     4681 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag
     4741 cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc
     4801 cccatctctg aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc
     4861 actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc
     4921 actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac
     4981 aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg
     5041 atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg
     5101 ctgccctctg gttatgtgtg ggagggctaa gcacttcatc gataaaataa aagattttat
     5161 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt
     5221 aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat
     5281 caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca
     5341 gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga
     5401 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
     5461 ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
     5521 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
     5581 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc
     5641 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
     5701 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
     5761 gcgggggtct ttcatttccg acttgtggtc tcgctgcctt gggagggtct cctctgagtg
     5821 attgactacc cgtcagcggg ggtcttcaca tgcagcatgt atcaaaatta atttggtttt
     5881 ttttcttaag tatttacatt aaatggccat agttgcatta atgaatcggc caacgcgcgg
     5941 ggagaggcgg tttgcgtatt ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6001 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6061 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6121 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6181 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6241 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6301 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6361 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6421 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6481 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6541 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6601 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6661 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6721 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6781 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6841 ccttttaaat taaaaatgaa gtttgcggcc gcaaatcaat ctaaagtata tatgagtaaa
     6901 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6961 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7021 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7081 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7141 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7201 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7261 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7321 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7381 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7441 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7501 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     7561 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7621 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7681 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7741 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7801 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7861 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc ac
//
