LOCUS       sequence-47115-e        6958 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 6958)
  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 6958)
  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: pIC389
FEATURES             Location/Qualifiers
     source          1..6958
                     /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(3373..3393)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3378..3707
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3558..3693
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3722..3769
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3788..4186
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4239..4832
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5033..5050)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5204..5792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5284..5303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5970..6761,6762..6830))
                     /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     6593..6612
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6831..6935)
                     /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 cgagcgactt aatctctcag accttcaaat
     2281 ggttcttggt ctgagctaca gcacaaataa ctccataatg tctgaaaata ctgaaattga
     2341 tagcagtttg aatggtgtac gccccaagtt gggacgccgg catagtatgg aaaatatgga
     2401 acttatgaag ttaacaccag aaaaggtaca gaactggaac agtgaaattc ttgctaagca
     2461 aaaacctctt attgccaaac cttctgcaaa gctactcttt gtcaacagac tgaaggggaa
     2521 aaaatacaaa acgaattcct ccactaaagt tctccaagat gccagtaatt ctattgacca
     2581 ccgaattcca aattctcaga ggaaaatcag tgcagatact attggagatg aagggttctt
     2641 tgacttatta agccgatttc aaagcaatag gatggatgat cagagatgtt gcttacaaga
     2701 aaagaactgc catacagctt caacaacaac ttcttccact ccccctaaaa tgatgctaaa
     2761 aacatcatct gttcctgtgg tatcccccaa cacggatgag tttttagatc ttcttgccag
     2821 ctcacagagt cgccgtctgg atgaccagag ggctagtttc agtaatttgc cagggcttcg
     2881 tctaacacaa aacagccagt cggtacttag ccacctgatg actaatgaca acaaagaggc
     2941 tgatgaagat ttctttgaca tccttgtaaa atgtcaagga tccagattag atgatcaaag
     3001 atgtgctcca ccacctgcta ccacaaaggg tccgacagta ccagatgaag actttttcag
     3061 ccttatttta cggtcccagg gaaagagaat ggatgaacag agagttcttt tacaaagaga
     3121 tcaaaacaga gacactgact ttgggctaaa ggactttttg caaaataatg ctttgttgga
     3181 gtttaaaaat tcagggaaaa aatcggcaga ccattagccg cgggcccggg atccaccgga
     3241 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     3301 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattcg ccagcacagt
     3361 ggtcgaccct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3421 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3481 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3541 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3601 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3661 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttgagcacg
     3721 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     3781 ctaaaccatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc
     3841 tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag
     3901 cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga
     3961 actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc
     4021 gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct
     4081 cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt
     4141 tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaagcac ttcatcgata
     4201 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
     4261 tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac ataactgaga
     4321 atagagaagt tcagatcaag gtcaggaaca gatggaacag ctgaatatgg gccaaacagg
     4381 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa
     4441 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     4501 gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca
     4561 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     4621 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
     4681 actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac
     4741 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt
     4801 gattgactac ccgtcagcgg gggtctttca tttccgactt gtggtctcgc tgccttggga
     4861 gggtctcctc tgagtgattg actacccgtc agcgggggtc ttcacatgca gcatgtatca
     4921 aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagtt gcattaatga
     4981 atcggccaac gcgcggggag aggcggtttg cgtattggcg ctcttccgct tcctcgctca
     5041 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5101 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5161 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5221 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5281 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5341 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5401 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5461 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5521 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5581 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5641 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5701 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5761 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5821 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5881 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt gcggccgcaa atcaatctaa
     5941 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6001 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6061 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6121 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6181 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6241 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6301 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6361 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6421 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6481 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6541 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc
     6601 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6661 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6721 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6781 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6841 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6901 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccac
//
