LOCUS       sequence-9784-ex        7169 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 7169)
  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 7169)
  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: pmEGFP_a_tubulin_IRES_puro2b
FEATURES             Location/Qualifiers
     source          1..7169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          74..303
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     286..305
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(361..935,936..938,939..947)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(528..545)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     755..774
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             967..1566
                     /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(967..986)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1463..1483
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(1702..1719)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1708..1932
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1979..1998
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2058..2074)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2058..2074)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2071..2093)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2082..2098
                     /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(2106..2112,2113..2130,2131..2136))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2151..2172
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2289..2306)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2460..3048)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2540..2559)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3219..4010,4011..4079))
                     /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     3842..3861
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4080..4184)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4259..4278)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4450..4829
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4830..5033
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4984..5004
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      5072..5081
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5078..5080,5081..5083,5084..5794)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5123..5144)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5384..5403)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 gatccgcggc cgcatagata actgatccag tgtgctggaa ttaattcgct gtctgcgagg
       61 gccagctgtt ggggtgagta ctccctctca aaagcgggca tgacttctgc gctaagattg
      121 tcagtttcca aaaacgagga ggatttgata ttcacctggc ccgcggtgat gcctttgagg
      181 gtggccgcgt ccatctggtc agaaaagaca atctttttgt tgtcaagctt gaggtgtggc
      241 aggcttgaga tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca
      301 caggtgtcca ctcccaggtc caactgcagg tcgagcatgc atctagggcg gccaattccg
      361 cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
      421 tgcgtttgtc tatatgtgat tttccaccat attgccgtct tttggcaatg tgagggcccg
      481 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
      541 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
      601 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
      661 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
      721 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
      781 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
      841 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
      901 ggacgtggtt ttcctttgaa aaacacgatg ataagcttgc cacaacccac aaggagacga
      961 ccttccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg
     1021 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac
     1081 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     1141 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     1201 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     1261 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     1321 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     1381 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     1441 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     1501 accgccgacg tcgagtgccc gaaggaccgc gcgacctggt gcatgacccg caagcccggt
     1561 gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatggc
     1621 tccgaccgaa gccgacccgg gcggccccgc cgaccccgca cccgcccccg aggcccaccg
     1681 actctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     1741 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     1801 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     1861 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     1921 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctcgagtgca ttctagttgt
     1981 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     2041 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2101 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2161 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2221 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2281 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2341 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2401 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2461 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2521 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2581 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2641 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2701 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2761 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2821 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2881 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2941 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3001 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3061 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3121 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3181 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3241 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3301 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3361 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3421 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3481 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3541 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3601 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3661 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3721 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3781 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3841 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3901 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3961 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4021 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4081 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4141 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4201 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta
     4261 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     4321 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     4381 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
     4441 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     4501 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     4561 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     4621 caatagggac tttccattga cgtcaatggg tggactattt acggtaaact gcccacttgg
     4681 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     4741 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     4801 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     4861 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     4921 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     4981 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
     5041 tgaaccgtca gatccgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct
     5101 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     5161 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     5221 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     5281 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     5341 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     5401 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     5461 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     5521 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     5581 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     5641 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     5701 gagcaaagac cccaacgaga agcgcgatca catggtcctt aaggagttcg tgaccgccgc
     5761 cgggatcact ctcggcatgg acgagctgta caagtccgga ctcagatctc gagtgcgtga
     5821 gtgcatctcc atccacgttg gccaggctgg tgtccagatt ggcaatgcct gctgggagct
     5881 ctactgcctg gaacacggca tccagcccga tggccagatg ccaagtgaca agaccattgg
     5941 gggaggagat gactccttca acaccttctt cagtgagacg ggcgctggca agcacgtgcc
     6001 ccgggctgtg tttgtagact tggaacccac agtcattgat gaagttcgca ctggcaccta
     6061 ccgccagctc ttccaccctg agcagctcat cacaggcaag gaagatgctg ccaataacta
     6121 tgcccgaggg cactacacca ttggcaagga gatcattgac cttgtgttgg accgaattcg
     6181 caagctggct gaccagtgca ccggtcttca gggcttcttg gttttccaca gctttggtgg
     6241 gggaactggt tctgggttca cctccctgct catggaacgt ctctcagttg attatggcaa
     6301 gaagtccaag ctggagttct ccatttaccc agcaccccag gtttccacag ctgtagttga
     6361 gccctacaac tccatcctca ccacccacac caccctggag cactctgatt gtgccttcat
     6421 ggtagacaat gaggccatct atgacatctg tcgtagaaac ctcgatatcg agcgcccaac
     6481 ctacactaac cttaaccgcc ttattagcca gattgtgtcc tccatcactg cttccctgag
     6541 atttgatgga gccctgaatg ttgacctgac agaattccag accaacctgg tgccctaccc
     6601 ccgcatccac ttccctctgg ccacatatgc ccctgtcatc tctgctgaga aagcctacca
     6661 tgaacagctt tctgtagcag agatcaccaa tgcttgcttt gagccagcca accagatggt
     6721 gaaatgtgac cctcgccatg gtaaatacat ggcttgctgc ctgttgtacc gtggtgacgt
     6781 ggttcccaaa gatgtcaatg ctgccattgc caccatcaaa accaagcgca gcatccagtt
     6841 tgtggattgg tgccccactg gcttcaaggt tggcatcaac taccagcctc ccactgtggt
     6901 gcctggtgga gacctggcca aggtacagag agctgtgtgc atgctgagca acaccacagc
     6961 cattgctgag gcctgggctc gcctggacca caagtttgac ctgatgtatg ccaagcgtgc
     7021 ctttgttcac tggtacgtgg gtgaggggat ggaggaaggc gagttttcag aggcccgtga
     7081 agatatggct gcccttgaga aggattatga ggaggttggt gtggattctg ttgaaggaga
     7141 gggtgaggaa gaaggagagg aatactaag
//
