LOCUS       sequence-62984-e        7870 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 7870)
  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 7870)
  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: Cenp-B-targeted Aurora B FRET sensor
FEATURES             Location/Qualifiers
     source          1..7870
                     /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      903..912
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1422..1431
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1428..2135
                     /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(2694..2696,2697..3410)
                     /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(2736..2757)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2997..3016)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     promoter        complement(3426..3444)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3427..3444)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3464..3481)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3470..3694
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3740..4168
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3827..3846)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4037..4058
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4177..4197)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4182..4511
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4362..4497
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4424..4443
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4578..5372
                     /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(4632..4651)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5242..5261
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5546..5667
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5583..5602)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5637..5656
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5716..5732)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5716..5732)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5729..5751)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5740..5756
                     /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(5764..5770,5771..5788,5789..5794))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5809..5830
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5947..5964)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6118..6703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6198..6217)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6874..7665,7666..7734))
                     /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     7497..7516
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7735..7839)
                     /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 ccgccaccat gggccccaag aggcgacagc tgacgttccg ggagaagtca cggatcatcc
      961 aggaggtgga ggagaatccg gacctgcgca agggcgagat cgcgcggcgc ttcaacatcc
     1021 cgccgtccac gctgagcacg atcctgaaga acaagcgcgc catcctggcg tcggagcgca
     1081 agtacggggt ggcctccacc tgccgcaaga ccaacaagct gtctccctac gacaagctcg
     1141 agggcttgct catcgcctgg ttccagcaga tccgcgccgc cggcctgccg gtcaagggca
     1201 tcatcctcaa ggagaaggcg ctgcgcatag ccgaggagct gggcatggac gacttcaccg
     1261 cctccaacgg ctggctggac cgcttccgcc ggcgccacgg cgtggtgtcc tgcagcggcg
     1321 tggcccgcgc ccgcgcgcga aacgctgccc cccgcacccc ggcggcgcct gccagtccgg
     1381 ccgcggtgcc ctcggagggc agtggcgggc tagcctcgag cgccaccatg gtgagcaagg
     1441 gcgaggagac cacaatgggc gtaatcaagc ccgacatgaa gatcaagctg aagatggagg
     1501 gcaacgtgaa tggccacgcc ttcgtgatcg agggcgaggg cgagggcaag ccctacgacg
     1561 gcaccaacac catcaacctg gaggtgaagg agggagcccc cctgcccttc tcctacgaca
     1621 ttctgaccac cgcgttcgcc tacggcaaca gggccttcac caagtacccc gacgacatcc
     1681 ccaactactt caagcagtcc ttccccgagg gctactcttg ggagcgcacc atgaccttcg
     1741 aggacaaggg catcgtgaag gtgaagtccg acatctccat ggaggaggac tccttcatct
     1801 acgagataca cctcaagggc gagaacttcc cccccaacgg ccccgtgatg cagaagaaga
     1861 ccaccggctg ggacgcctcc accgagagga tgtacgtgcg cgacggcgtg ctgaagggcg
     1921 acgtcaagca caagctgctg ctggagggcg gcggccacca ccgcgttgac ttcaagacca
     1981 tctacagggc caagaaggcg gtgaagctgc ccgactatca ctttgtggac caccgcatcg
     2041 agatcctgaa ccacgacaag gactacaaca aggtgaccgt ttacgagagc gccgtggccc
     2101 gcaactccac cgacggcatg gacgagctgt acaaggctgg taccaagggt aatggtaggt
     2161 ttttaacttt aaaaccattg cctgacagca ttattcaaga aagcctggag attcagcaag
     2221 gtgtgaatcc atttttcatt ggtagatccg aggattgcaa ttgtaaaatt gaagacagta
     2281 ggttgtctcg agttcattgc ttcattttca aaaagaggca tgctgtaggc aaaagcatgt
     2341 atgaatctcc ggcacaaggt ttagatgata tttggtattg ccacaccgga actaacgtga
     2401 gctatttaaa taataaccgc atgatacagg gtacgaaatt ccttttacaa gacggagatg
     2461 aaatcaagat catttgggat aaaaacaata aatttgtcat tggctttaaa gtggaaatta
     2521 acgatactac aggtctgttt aacgagggat taggtatgtt acaagaacaa agagtagtac
     2581 ttaagcaaac agccgaagaa aaagatttgg tgaaaaagtt aggaggatcc ggcggcaagg
     2641 tgaacaagat cgtgaagaac cgccgcaccg tggccatcgg tggttccgga ggagtgagca
     2701 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     2761 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     2821 ccctgaagct gctctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2881 ccctgggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     2941 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     3001 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     3061 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     3121 acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     3181 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
     3241 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     3301 accagtccgc cctgttcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     3361 tcctgaccgc cgccgggatc actgaaggca tgaacgagct gtacaagtaa tctagagggc
     3421 cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct
     3481 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3541 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3601 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3661 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3721 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3781 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3841 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3901 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3961 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4021 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4081 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4141 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     4201 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4261 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4321 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4381 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4441 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4501 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     4561 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4621 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4681 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4741 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4801 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4861 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4921 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4981 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5041 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5101 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5161 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5221 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5281 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5341 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5401 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5461 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5521 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5581 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5641 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5701 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5761 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5821 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5881 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5941 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6001 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6061 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6121 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6181 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6241 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6301 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6361 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6421 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6481 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6541 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6601 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     6661 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6721 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6781 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6841 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6901 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6961 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7021 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7081 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7141 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7201 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7261 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7321 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7381 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7441 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7501 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7561 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7621 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7681 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7741 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7801 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7861 acctgacgtc
//
