LOCUS       sequence-53370-e        7062 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 7062)
  AUTHORS   Wurzenberger C, Held M, Lampson MA, Poser I, Hyman AA, Gerlich DW
  TITLE     Sds22 and Repo-Man stabilize chromosome segregation by counteracting
            Aurora B on anaphase kinetochores.
  JOURNAL   J Cell Biol. 2012 Jul 23;198(2):173-83. Epub 2012 Jul 16.
  PUBMED    22801782
REFERENCE   2  (bases 1 to 7062)
  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: "J Cell 
            Biol."; date: "2012-07-23"; volume: "198(2)"; pages: "173-83. Epub 
            2012 Jul 16"
COMMENT     Sequence Label: pCyPet-FHA2-AuroraB Substrate-YPet-IRES-puro2b
FEATURES             Location/Qualifiers
     source          1..7062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          80..309
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     292..311
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(367..941,942..944,945..953)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(534..551)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     761..780
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             973..1572
                     /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(973..992)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1469..1489
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(1708..1725)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1714..1938
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1985..2004
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2064..2080)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2064..2080)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2077..2099)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2088..2104
                     /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(2112..2118,2119..2136,2137..2142))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2157..2178
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2295..2312)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2466..3054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2546..2565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3225..4016,4017..4085))
                     /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     3848..3867
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4086..4190)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4265..4284)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4456..4835
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4836..5039
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4990..5010
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      5075..5084
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5081..5083,5084..5086,5087..5797)
                     /codon_start=1
                     /product="monomeric super enhanced variant of CFP (Matsuda 
                     et al., 2008)"
                     /label=mseCFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFGGIVPILVELEGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSVMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKARHNITDGSVQLADHYQQNTPIGDGPVILPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             join(5081..5083,5084..5086,5087..5797)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFGGIVPILVELEGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSVMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKARHNITDGSVQLADHYQQNTPIGDGPVILPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             join(5081..5083,5084..5086,5087..5797)
                     /codon_start=1
                     /product="enhanced CFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFGGIVPILVELEGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSVMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKARHNITDGSVQLADHYQQNTPIGDGPVILPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             join(6356..6358,6359..6871)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LLCTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIE"
ORIGIN
        1 gtacaagtaa agcggccgca tagataactg atccagtgtg ctggaattaa ttcgctgtct
       61 gcgagggcca gctgttgggg tgagtactcc ctctcaaaag cgggcatgac ttctgcgcta
      121 agattgtcag tttccaaaaa cgaggaggat ttgatattca cctggcccgc ggtgatgcct
      181 ttgagggtgg ccgcgtccat ctggtcagaa aagacaatct ttttgttgtc aagcttgagg
      241 tgtggcaggc ttgagatctg gccatacact tgagtgacaa tgacatccac tttgcctttc
      301 tctccacagg tgtccactcc caggtccaac tgcaggtcga gcatgcatct agggcggcca
      361 attccgcccc tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg
      421 ccggtgtgcg tttgtctata tgtgattttc caccatattg ccgtcttttg gcaatgtgag
      481 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
      541 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
      601 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
      661 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
      721 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt
      781 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc
      841 tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa
      901 ccacggggac gtggttttcc tttgaaaaac acgatgataa gcttgccaca acccacaagg
      961 agacgacctt ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     1021 ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     1081 gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     1141 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     1201 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     1261 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     1321 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     1381 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     1441 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     1501 accgtcaccg ccgacgtcga gtgcccgaag gaccgcgcga cctggtgcat gacccgcaag
     1561 cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc
     1621 catggctccg accgaagccg acccgggcgg ccccgccgac cccgcacccg cccccgaggc
     1681 ccaccgactc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     1741 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     1801 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     1861 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     1921 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctcg agtgcattct
     1981 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     2041 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2101 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     2161 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2221 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2281 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2341 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2401 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2461 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2521 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2581 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2641 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2701 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2761 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2821 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2881 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     2941 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3001 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3061 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3121 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3181 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3241 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3301 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3361 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3421 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3481 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3541 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3601 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3661 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3721 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3781 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3841 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3901 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3961 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4021 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4081 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4141 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4201 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc
     4261 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     4321 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     4381 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
     4441 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
     4501 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     4561 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     4621 taacgccaat agggactttc cattgacgtc aatgggtgga ctatttacgg taaactgccc
     4681 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     4741 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     4801 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     4861 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     4921 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     4981 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     5041 gtttagtgaa ccgtcagatc cgctagcaag cttcgccacc atggtgagca agggcgagga
     5101 gctgttcggc gggatcgtgc ccatcctggt cgagctggag ggcgacgtaa acggccacag
     5161 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     5221 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgacctg
     5281 gggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     5341 cgtcatgccc gaaggctacg tccaggagcg taccatcttc ttcaaggacg acggcaacta
     5401 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     5461 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacat
     5521 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg cccacttcaa
     5581 ggcccgccac aacatcacgg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     5641 ccccatcggc gacggccccg tgatcctgcc cgacaaccac tacctgagca cccagtccaa
     5701 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     5761 cgccgggatc actcacggca tggacgagct gtacaaggct ggtaccaagg gtaatggtag
     5821 gtttttaact ttaaaaccat tgcctgacag cattattcaa gaaagcctgg agattcagca
     5881 aggtgtgaat ccatttttca ttggtagatc cgaggattgc aattgtaaaa ttgaagacag
     5941 taggttgtct cgagttcatt gcttcatttt caaaaagagg catgctgtag gcaaaagcat
     6001 gtatgaatct ccggcacaag gtttagatga tatttggtat tgccacaccg gaactaacgt
     6061 gagctattta aataataacc gcatgataca gggtacgaaa ttccttttac aagacggaga
     6121 tgaaatcaag atcatttggg ataaaaacaa taaatttgtc attggcttta aagtggaaat
     6181 taacgatact acaggtctgt ttaacgaggg attaggtatg ttacaagaac aaagagtagt
     6241 acttaagcaa acagccgaag aaaaagattt ggtgaaaaag ttaggaggat ccggcggcaa
     6301 ggtgaacaag atcgtgaaga accgccgcac cgtggccatc ggtggttccg gaggagtgag
     6361 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     6421 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     6481 gaccctgaag ctgctctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     6541 caccctgggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     6601 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     6661 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     6721 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     6781 gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     6841 ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta
     6901 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     6961 ctaccagtcc gccctgttca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     7021 gttcctgacc gccgccggga tcactgaagg catgaacgag ct
//
