LOCUS       sequence-62980-e        7984 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 7984)
  AUTHORS   Liu D, Vader G, Vromans MJ, Lampson MA, Lens SM
  TITLE     Sensing chromosome bi-orientation by spatial separation of aurora B 
            kinase from kinetochore substrates.
  JOURNAL   Science. 2009 Mar 6;323(5919):1350-3. doi: 10.1126/science.1167000. 
            Epub 2009 Jan 15.
  PUBMED    19150808
REFERENCE   2  (bases 1 to 7984)
  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: 
            "10.1126/science.1167000"; journalName: "Science"; date: "2009-03-6-
            6"; volume: "323"; issue: "5919"; pages: "1350-3"
COMMENT     Sequence Label: nonphosphorylatable Mis12-targeted Aurora B FR...
FEATURES             Location/Qualifiers
     source          1..7984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1536..1545
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1542..2249
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     CDS             join(2808..2810,2811..3524)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LLCTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLLE
                     FLTAAGITEGMNELYK"
     primer_bind     complement(2850..2871)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3111..3130)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     promoter        complement(3540..3558)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3541..3558)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3578..3595)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3584..3808
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3854..4282
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3941..3960)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4151..4172
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4291..4311)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4296..4625
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4476..4611
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4538..4557
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4692..5486
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4746..4765)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5356..5375
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5660..5781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5697..5716)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5751..5770
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5830..5846)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5830..5846)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5843..5865)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5854..5870
                     /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(5878..5884,5885..5902,5903..5908))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5923..5944
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6061..6078)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6232..6817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6312..6331)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6988..7779,7780..7848))
                     /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     7611..7630
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7849..7953)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggcgcg
      901 ccaccatgtc tgtggatcca atgacctacg aggcccagtt ctttggcttc acgccacaaa
      961 cgtgcatgct tcggatctac attgcatttc aagactacct atttgaagtg atgcaggccg
     1021 ttgaacaggt tattctgaag aagctggatg gcatcccaga ctgtgacatt agcccagtgc
     1081 agattcgcaa atgcacagag aagtttcttt gcttcatgaa aggacatttt gataaccttt
     1141 ttagcaaaat ggagcaactg tttttgcagc tgattttacg tattccctca aacatcttgc
     1201 ttcctgaaga taaatgtaag gagacacctt atagtgagga agattttcag catctccaga
     1261 aagaaattga acagttacag gagaagtaca agactgaatt atgtactaag caggcccttc
     1321 ttgcagaatt agaagagcaa aaaattgttc aggccaaact caaacagacg ttgactttct
     1381 ttgatgagct tcataatgtt ggcagagatc atgggactag tgattttagg gagagtttag
     1441 tatccctggt tcagaactcc agaaaactac agaacattag agacaatgtg gaaaaggaat
     1501 cgaaacgact gaaaatatct atgctagcct cgagcgccac catggtgagc aagggcgagg
     1561 agaccacaat gggcgtaatc aagcccgaca tgaagatcaa gctgaagatg gagggcaacg
     1621 tgaatggcca cgccttcgtg atcgagggcg agggcgaggg caagccctac gacggcacca
     1681 acaccatcaa cctggaggtg aaggagggag cccccctgcc cttctcctac gacattctga
     1741 ccaccgcgtt cgcctacggc aacagggcct tcaccaagta ccccgacgac atccccaact
     1801 acttcaagca gtccttcccc gagggctact cttgggagcg caccatgacc ttcgaggaca
     1861 agggcatcgt gaaggtgaag tccgacatct ccatggagga ggactccttc atctacgaga
     1921 tacacctcaa gggcgagaac ttccccccca acggccccgt gatgcagaag aagaccaccg
     1981 gctgggacgc ctccaccgag aggatgtacg tgcgcgacgg cgtgctgaag ggcgacgtca
     2041 agcacaagct gctgctggag ggcggcggcc accaccgcgt tgacttcaag accatctaca
     2101 gggccaagaa ggcggtgaag ctgcccgact atcactttgt ggaccaccgc atcgagatcc
     2161 tgaaccacga caaggactac aacaaggtga ccgtttacga gagcgccgtg gcccgcaact
     2221 ccaccgacgg catggacgag ctgtacaagg ctggtaccaa gggtaatggt aggtttttaa
     2281 ctttaaaacc attgcctgac agcattattc aagaaagcct ggagattcag caaggtgtga
     2341 atccattttt cattggtaga tccgaggatt gcaattgtaa aattgaagac agtaggttgt
     2401 ctcgagttca ttgcttcatt ttcaaaaaga ggcatgctgt aggcaaaagc atgtatgaat
     2461 ctccggcaca aggtttagat gatatttggt attgccacac cggaactaac gtgagctatt
     2521 taaataataa ccgcatgata cagggtacga aattcctttt acaagacgga gatgaaatca
     2581 agatcatttg ggataaaaac aataaatttg tcattggctt taaagtggaa attaacgata
     2641 ctacaggtct gtttaacgag ggattaggta tgttacaaga acaaagagta gtacttaagc
     2701 aaacagccga agaaaaagat ttggtgaaaa agttaggagg atccggcggc aaggtgaaca
     2761 agatcgtgaa gaaccgccgc gccgtggcca tcggtggctc cggaggagtg agcaagggcg
     2821 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2881 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2941 agctgctctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg
     3001 gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     3061 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     3121 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3181 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3241 acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     3301 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     3361 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     3421 ccgccctgtt caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcctga
     3481 ccgccgccgg gatcactgaa ggcatgaacg agctgtacaa gtaatctaga gggccctatt
     3541 ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     3601 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3661 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3721 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3781 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3841 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3901 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3961 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4021 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4081 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4141 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4201 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4261 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4321 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4381 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4441 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4501 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4561 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4621 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4681 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4741 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4801 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4861 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4921 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4981 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5041 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5101 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5161 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5221 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5281 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5341 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5401 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5461 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5521 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5581 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5641 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5701 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5761 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5821 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5881 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5941 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6001 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6061 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6121 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6181 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6241 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6301 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6361 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6421 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6481 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6541 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6601 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6661 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6721 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     6781 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6841 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6901 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6961 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7021 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7081 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7141 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7201 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7261 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7321 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7381 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7441 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7501 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7561 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7621 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7681 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7741 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7801 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7861 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7921 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7981 cgtc
//
