LOCUS       sequence-466944-        6239 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 6239)
  AUTHORS   Steigemann P, Wurzenberger C, Schmitz MH, Held M, Guizetti J, Maar 
            S, Gerlich DW
  TITLE     Aurora B-mediated abscission checkpoint protects against 
            tetraploidization.
  JOURNAL   Cell. 2009 Feb 6. 136(3):473-84.
  PUBMED    19203582
REFERENCE   2  (bases 1 to 6239)
  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"; journalName: "Cell. 2009 
            Feb 6. 136(3):473-84."
COMMENT     Sequence Label: pH2B_mCherry_IRES_puro2
FEATURES             Location/Qualifiers
     source          1..6239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             293..670
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      683..692
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             689..1399
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(834..852)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1102..1121
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          1469..1698
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1681..1700
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(1771..2345,2346..2348,2349..2357)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1938..1955)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2165..2184
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2377..2976
                     /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(2377..2396)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2873..2893
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3112..3129)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3118..3342
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3389..3408
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3468..3484)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3468..3484)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3481..3503)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3492..3508
                     /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(3516..3522,3523..3540,3541..3546))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3561..3582
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3699..3716)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3870..4458)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3950..3969)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4629..5420,5421..5489))
                     /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     5252..5271
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5490..5594)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5669..5688)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5860..6239
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct cgagctcaag cttcgaattc tgcagtcgac ggtaccgcca ccatgccaga
      301 gccagcgaag tctgctcccg ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca
      361 gaagaaaggc ggcaagaagc gcaagcgcag ccgcaaggag agctattcca tctatgtgta
      421 caaggttctg aagcaggtcc accctgacac cggcatttcg tccaaggcca tgggcatcat
      481 gaattcgttt gtgaacgaca ttttcgagcg catcgcaggt gaggcttccc gcctggcgca
      541 ttacaacaag cgctcgacca tcacctccag ggagatccag acggccgtgc gcctgctgct
      601 gcctggggag ttggccaagc acgccgtgtc cgagggtact aaggccatca ccaagtacac
      661 cagcgctaag gatccaccgg tcgccaccat ggtgagcaag ggcgaggagg ataacatggc
      721 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
      781 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
      841 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
      901 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
      961 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     1021 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     1081 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     1141 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     1201 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     1261 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     1321 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     1381 ggacgagctg tacaagtaaa gcggccgcat agataactga tccagtgtgc tggaattaat
     1441 tcgctgtctg cgagggccag ctgttggggt gagtactccc tctcaaaagc gggcatgact
     1501 tctgcgctaa gattgtcagt ttccaaaaac gaggaggatt tgatattcac ctggcccgcg
     1561 gtgatgcctt tgagggtggc cgcgtccatc tggtcagaaa agacaatctt tttgttgtca
     1621 agcttgaggt gtggcaggct tgagatctgg ccatacactt gagtgacaat gacatccact
     1681 ttgcctttct ctccacaggt gtccactccc aggtccaact gcaggtcgag catgcatcta
     1741 gggcggccaa ttccgcccct ctcccccccc cccctctccc tccccccccc ctaacgttac
     1801 tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat
     1861 attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat
     1921 tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga
     1981 agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca
     2041 gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac
     2101 acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt
     2161 caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca
     2221 ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt
     2281 aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg
     2341 ataagcttgc cacaacccac aaggagacga ccttccatga ccgagtacaa gcccacggtg
     2401 cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc
     2461 gactaccccg ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag
     2521 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     2581 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     2641 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     2701 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     2761 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     2821 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     2881 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     2941 cgcacctggt gcatgacccg caagcccggt gcctgacgcc cgccccacga cccgcagcgc
     3001 ccgaccgaaa ggagcgcacg accccatggc tccgaccgaa gccacccggg gcggccccgc
     3061 cgaccccgca cccgcccccg aggcccaccg actctagagc tcgctgatca gcctcgactg
     3121 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3181 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3241 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3301 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3361 ccagctgggg ctcgagtgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     3421 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     3481 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3541 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3601 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3661 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3721 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3781 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3841 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3901 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3961 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4021 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4081 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4141 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4201 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4261 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4321 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4381 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4441 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4501 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4561 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4621 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4681 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4741 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4801 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4861 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4921 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4981 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5041 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5101 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5161 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5221 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5281 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5341 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5401 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5461 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5521 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5581 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     5641 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5701 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     5761 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     5821 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
     5881 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     5941 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     6001 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     6061 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     6121 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     6181 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatg
//
