LOCUS       sequence-9783-ex        7136 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 7136)
  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 7136)
  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: pmRFP_a_tubulin_IRES_puro2b
FEATURES             Location/Qualifiers
     source          1..7136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1287
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             613..1287
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(743..761)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1011..1030
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1205..1228
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     intron          2745..2974
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2957..2976
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3032..3606,3607..3609,3610..3618)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3199..3216)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3426..3445
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3638..4237
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(3638..3657)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4134..4154
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(4373..4390)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4379..4603
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4650..4669
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4729..4745)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4729..4745)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4742..4764)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4753..4769
                     /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(4777..4783,4784..4801,4802..4807))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4822..4843
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4960..4977)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5131..5719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5211..5230)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5890..6681,6682..6750))
                     /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     6513..6532
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6751..6855)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6930..6949)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(7121..7136,1..364)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggcctc ctccgaggac gtcatcaagg agttcatgcg cttcaaggtg
      661 cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
      721 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc
      781 gcctgggaca tcctgtcccc tcagttccag tacggctcca aggcctacgt gaagcacccc
      841 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
      901 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
      961 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     1021 cagaagaaga ccatgggctg ggaggcctcc accgagcgga tgtaccccga ggacggcgcc
     1081 ctgaagggcg agatcaagat gaggctgaag ctgaaggacg gcggccacta cgacgccgag
     1141 gtcaagacca cctacatggc caagaagccc gtgcagctgc ccggcgccta caagaccgac
     1201 atcaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc
     1261 gccgagggcc gccactccac cggcgccctg tacaagtccg gactcagatc tcgagtgcgt
     1321 gagtgcatct ccatccacgt tggccaggct ggtgtccaga ttggcaatgc ctgctgggag
     1381 ctctactgcc tggaacacgg catccagccc gatggccaga tgccaagtga caagaccatt
     1441 gggggaggag atgactcctt caacaccttc ttcagtgaga cgggcgctgg caagcacgtg
     1501 ccccgggctg tgtttgtaga cttggaaccc acagtcattg atgaagttcg cactggcacc
     1561 taccgccagc tcttccaccc tgagcagctc atcacaggca aggaagatgc tgccaataac
     1621 tatgcccgag ggcactacac cattggcaag gagatcattg accttgtgtt ggaccgaatt
     1681 cgcaagctgg ctgaccagtg caccggtctt cagggcttct tggttttcca cagctttggt
     1741 gggggaactg gttctgggtt cacctccctg ctcatggaac gtctctcagt tgattatggc
     1801 aagaagtcca agctggagtt ctccatttac ccagcacccc aggtttccac agctgtagtt
     1861 gagccctaca actccatcct caccacccac accaccctgg agcactctga ttgtgccttc
     1921 atggtagaca atgaggccat ctatgacatc tgtcgtagaa acctcgatat cgagcgccca
     1981 acctacacta accttaaccg ccttattagc cagattgtgt cctccatcac tgcttccctg
     2041 agatttgatg gagccctgaa tgttgacctg acagaattcc agaccaacct ggtgccctac
     2101 ccccgcatcc acttccctct ggccacatat gcccctgtca tctctgctga gaaagcctac
     2161 catgaacagc tttctgtagc agagatcacc aatgcttgct ttgagccagc caaccagatg
     2221 gtgaaatgtg accctcgcca tggtaaatac atggcttgct gcctgttgta ccgtggtgac
     2281 gtggttccca aagatgtcaa tgctgccatt gccaccatca aaaccaagcg cagcatccag
     2341 tttgtggatt ggtgccccac tggcttcaag gttggcatca actaccagcc tcccactgtg
     2401 gtgcctggtg gagacctggc caaggtacag agagctgtgt gcatgctgag caacaccaca
     2461 gccattgctg aggcctgggc tcgcctggac cacaagtttg acctgatgta tgccaagcgt
     2521 gcctttgttc actggtacgt gggtgagggg atggaggaag gcgagttttc agaggcccgt
     2581 gaagatatgg ctgcccttga gaaggattat gaggaggttg gtgtggattc tgttgaagga
     2641 gagggtgagg aagaaggaga ggaatactaa ggatccgcgg ccgcatagat aactgatcca
     2701 gtgtgctgga attaattcgc tgtctgcgag ggccagctgt tggggtgagt actccctctc
     2761 aaaagcgggc atgacttctg cgctaagatt gtcagtttcc aaaaacgagg aggatttgat
     2821 attcacctgg cccgcggtga tgcctttgag ggtggccgcg tccatctggt cagaaaagac
     2881 aatctttttg ttgtcaagct tgaggtgtgg caggcttgag atctggccat acacttgagt
     2941 gacaatgaca tccactttgc ctttctctcc acaggtgtcc actcccaggt ccaactgcag
     3001 gtcgagcatg catctagggc ggccaattcc gcccctctcc ctcccccccc cctaacgtta
     3061 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtga ttttccacca
     3121 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
     3181 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
     3241 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
     3301 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
     3361 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
     3421 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
     3481 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt
     3541 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
     3601 gataagcttg ccacaaccca caaggagacg accttccatg accgagtaca agcccacggt
     3661 gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc
     3721 cgactacccc gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga
     3781 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     3841 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     3901 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     3961 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     4021 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     4081 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     4141 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgagtgcc cgaaggaccg
     4201 cgcgacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg
     4261 cccgaccgaa aggagcgcac gaccccatgg ctccgaccga agccgacccg ggcggccccg
     4321 ccgaccccgc acccgccccc gaggcccacc gactctagag ctcgctgatc agcctcgact
     4381 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     4441 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     4501 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     4561 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     4621 accagctggg gctcgagtgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4681 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4741 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4801 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4861 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4921 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4981 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5041 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5101 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5161 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5221 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5281 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     5341 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5401 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5461 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5521 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5581 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5641 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5701 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5761 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5821 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5881 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5941 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6001 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6061 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6121 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6181 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6241 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6301 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6361 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6421 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6481 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6541 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6601 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6661 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6721 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6781 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6841 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     6901 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     6961 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     7021 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     7081 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgac
//
