LOCUS       sequence-383844-        5746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of DsRed fused to a positively charged polypeptide (2 x 
            KSG) . Overall charge on tetramer: +9e.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5746)
  AUTHORS   Schneider MWG, Gibson BA, Otsuka S, Spicer MFD, Petrovic M, Blaukopf
            C, Langer CCH, Batty P, Nagaraju T, Doolittle LK, Rosen MK, Gerlich 
            DW
  TITLE     A mitotic chromatin phase transition prevents perforation by 
            microtubules.
  JOURNAL   Nature. 2022 Sep;609(7925):183-190. doi: 10.1038/s41586-022-05027-y.
            Epub 2022 Aug 3.
  PUBMED    35922507
REFERENCE   2  (bases 1 to 5746)
  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: 
            "10.1038/s41586-022-05027-y"; journalName: "Nature"; date: 
            "2022-09"; volume: "609"; issue: "7925"; pages: "183-190"
COMMENT     Sequence Label: DsRed +9e
FEATURES             Location/Qualifiers
     source          1..5746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(64..855,856..924))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     687..706
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(925..1029)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1104..1123)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1295..1674
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1675..1878
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1829..1849
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1933..2607
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="RSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVKL
                     KVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHKA
                     LKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRHHL
                     FL"
     primer_bind     complement(2110..2130)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2328..2347
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2522..2545
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     intron          2651..2880
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2863..2882
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(2952..3526,3527..3529,3530..3538)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3119..3136)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3346..3365
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3558..4157
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3558..3577)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4054..4074
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(4293..4310)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4299..4523
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4570..4589
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4649..4665)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4649..4665)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4662..4684)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4673..4689
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4697..4703,4704..4721,4722..4727))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4742..4763
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4880..4897)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5051..5639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5131..5150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
       61 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
      121 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
      181 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
      241 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
      301 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
      361 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
      421 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
      481 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
      541 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
      601 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
      661 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
      721 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
      781 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
      841 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
      901 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
      961 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     1021 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     1081 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     1141 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     1201 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     1261 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
     1321 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     1381 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     1441 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     1501 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     1561 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     1621 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     1681 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     1741 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     1801 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     1861 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctaggccac catgaagagc
     1921 ggcaaaagcg gtcgctcctc caagaacgtc atcaaggagt tcatgcgctt caaggtgcgc
     1981 atggagggca ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     2041 tacgagggcc acaacaccgt gaagctgaag gtgaccaagg gcggccccct gcccttcgcc
     2101 tgggacatcc tgtcccccca gttccagtac ggctccaagg tgtacgtgaa gcaccccgcc
     2161 gacatccccg actacaagaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     2221 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggctgc
     2281 ttcatctaca aggtgaagtt catcggcgtg aacttcccct ccgacggccc cgtaatgcag
     2341 aagaagacca tgggctggga ggcctccacc gagcgcctgt acccccgcga cggcgtgctg
     2401 aagggcgaga tccacaaggc cctgaagctg aaggacggcg gccactacct ggtggagttc
     2461 aagtccatct acatggccaa gaagcccgtg cagctgcccg gctactacta cgtggactcc
     2521 aagctggaca tcacctccca caacgaggac tacaccatcg tggagcagta cgagcgcacc
     2581 gagggccgcc accacctgtt cctgtagtgt gctggaatta attcgctgtc tgcgagggcc
     2641 agctgttggg gtgagtactc cctctcaaaa gcgggcatga cttctgcgct aagattgtca
     2701 gtttccaaaa acgaggagga tttgatattc acctggcccg cggtgatgcc tttgagggtg
     2761 gccgcgtcca tctggtcaga aaagacaatc tttttgttgt caagcttgag gtgtggcagg
     2821 cttgagatct ggccatacac ttgagtgaca atgacatcca ctttgccttt ctctccacag
     2881 gtgtccactc ccaggtccaa ctgcaggtcg agcatgcatc tagggcggcc aattccgccc
     2941 ctctcccccc ccccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa
     3001 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     3061 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     3121 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     3181 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     3241 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     3301 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     3361 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     3421 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     3481 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataagcttg ccacaaccca
     3541 caaggagacg accttccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     3601 acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     3661 acaccgtcga cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     3721 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     3781 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     3841 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     3901 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     3961 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     4021 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     4081 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     4141 gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac
     4201 gaccccatgg ctccgaccga agccacccgg ggcggccccg ccgaccccgc acccgccccc
     4261 gaggcccacc gactctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     4321 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     4381 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     4441 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     4501 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctcgagtgc
     4561 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4621 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4681 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4741 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4801 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4861 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4921 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4981 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5041 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5101 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5161 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5221 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5281 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5341 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5401 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5461 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5521 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5581 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5641 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5701 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcct
//
