LOCUS       sequence-383843-        5746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of DsRed fused to a negatively charged polypeptide. 
            Overall charge on tetramer: -7e.
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 -7e
FEATURES             Location/Qualifiers
     source          1..5746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(65..856,857..925))
                     /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     688..707
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(926..1030)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1105..1124)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1296..1675
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1676..1879
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1830..1850
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1907..1916
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1934..2608
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="RSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVKL
                     KVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHKA
                     LKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRHHL
                     FL"
     primer_bind     complement(2111..2131)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2329..2348
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2523..2546
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     intron          2652..2881
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2864..2883
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(2953..3527,3528..3530,3531..3539)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3120..3137)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3347..3366
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3559..4158
                     /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(3559..3578)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4055..4075
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(4294..4311)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4300..4524
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4571..4590
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4650..4666)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4650..4666)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4663..4685)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4674..4690
                     /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(4698..4704,4705..4722,4723..4728))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4743..4764
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4881..4898)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5052..5640)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5132..5151)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
       61 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
      121 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
      181 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
      241 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
      301 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
      361 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
      421 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
      481 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
      541 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
      601 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
      661 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
      721 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
      781 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
      841 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
      901 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
      961 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1021 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     1081 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     1141 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     1201 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     1261 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
     1321 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     1381 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     1441 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     1501 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     1561 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     1621 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     1681 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     1741 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     1801 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     1861 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctaggcca ccatggagag
     1921 cggcgagagc ggtcgctcct ccaagaacgt catcaaggag ttcatgcgct tcaaggtgcg
     1981 catggagggc accgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     2041 ctacgagggc cacaacaccg tgaagctgaa ggtgaccaag ggcggccccc tgcccttcgc
     2101 ctgggacatc ctgtcccccc agttccagta cggctccaag gtgtacgtga agcaccccgc
     2161 cgacatcccc gactacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     2221 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggctg
     2281 cttcatctac aaggtgaagt tcatcggcgt gaacttcccc tccgacggcc ccgtaatgca
     2341 gaagaagacc atgggctggg aggcctccac cgagcgcctg tacccccgcg acggcgtgct
     2401 gaagggcgag atccacaagg ccctgaagct gaaggacggc ggccactacc tggtggagtt
     2461 caagtccatc tacatggcca agaagcccgt gcagctgccc ggctactact acgtggactc
     2521 caagctggac atcacctccc acaacgagga ctacaccatc gtggagcagt acgagcgcac
     2581 cgagggccgc caccacctgt tcctgtagtg tgctggaatt aattcgctgt ctgcgagggc
     2641 cagctgttgg ggtgagtact ccctctcaaa agcgggcatg acttctgcgc taagattgtc
     2701 agtttccaaa aacgaggagg atttgatatt cacctggccc gcggtgatgc ctttgagggt
     2761 ggccgcgtcc atctggtcag aaaagacaat ctttttgttg tcaagcttga ggtgtggcag
     2821 gcttgagatc tggccataca cttgagtgac aatgacatcc actttgcctt tctctccaca
     2881 ggtgtccact cccaggtcca actgcaggtc gagcatgcat ctagggcggc caattccgcc
     2941 cctctccccc cccccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga
     3001 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     3061 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     3121 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     3181 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     3241 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     3301 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     3361 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     3421 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     3481 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagctt gccacaaccc
     3541 acaaggagac gaccttccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     3601 gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     3661 cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     3721 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     3781 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     3841 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     3901 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     3961 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     4021 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     4081 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     4141 cgcaagcccg gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca
     4201 cgaccccatg gctccgaccg aagccacccg gggcggcccc gccgaccccg cacccgcccc
     4261 cgaggcccac cgactctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     4321 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     4381 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     4441 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     4501 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctcgagtg
     4561 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4621 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4681 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4741 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4801 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4861 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4921 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4981 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5041 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5101 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5161 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5221 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5281 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5341 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5401 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5461 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5521 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5581 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5641 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5701 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcc
//
