LOCUS       sequence-225044-        6928 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 6928)
  AUTHORS   Wordeman L, Wagenbach M, von Dassow G
  TITLE     MCAK facilitates chromosome movement by promoting kinetochore 
            microtubule turnover.
  JOURNAL   J Cell Biol. 2007 Dec 3. 179(5):869-79.
  PUBMED    18039936
REFERENCE   2  (bases 1 to 6928)
  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. 2007 Dec 3. 179(5):869-79."
COMMENT     Sequence Label: pMX215
FEATURES             Location/Qualifiers
     source          1..6928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     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"
     CDS             640..1314
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             640..1314
                     /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(770..788)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1038..1057
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1232..1255
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             3520..3537
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3716..3837
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3753..3772)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3807..3826
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3844..4299)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3981..4002)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4193..4212
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4326..4430
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4432..4789
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4432..4623
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4455..4475)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4640..4775
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4824..5618
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4824..5618
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4878..4897)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5488..5507
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5806..5825)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5850..5897
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6226..6814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6715..6734
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag 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 ccggactcag atctcgagct caagcttcga attcccacca tggcctcctc cgaggacgtc
      661 atcaaggagt tcatgcgctt caaggtgcgc atggagggct ccgtgaacgg ccacgagttc
      721 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
      781 gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccctca gttccagtac
      841 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
      901 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
      961 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     1021 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctccacc
     1081 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagatgag gctgaagctg
     1141 aaggacggcg gccactacga cgccgaggtc aagaccacct acatggccaa gaagcccgtg
     1201 cagctgcccg gcgcctacaa gaccgacatc aagctggaca tcacctccca caacgaggac
     1261 tacaccatcg tggaacagta cgagcgcgcc gagggccgcc actccaccgg cgccgatatc
     1321 tccggaatgg atccactagt ccagtgtggt ggaattgccc ttgccatgga gtcgcttccc
     1381 gctcgattgt ttcctggtct ctccatcaag attcaacgca gtaatggctt aattcacagt
     1441 gccaacataa gcacagtgaa cgtggagaaa tcctgtgttt cagtggaatg gatagaagga
     1501 ggtaacacga aaggcaaaga gattgatttt gatgatgtgg ctgcaataaa cccagaactt
     1561 ctacagcttc ttcccttaca cccaaaggac aatctgccac tgcaggaaaa tgtaacagtt
     1621 ccgaagcaaa aacgcagagc agtcaactcc aaaattccag ctccaaaaga aggtcttcga
     1681 gcacgtgcaa ctcgcatggc aactgtccca gaggttcgaa ttgccactca ggagaatgaa
     1741 atggaggtgg agctgcctgt ggctacaaac tctcgaaagc agttttcggt tgccactggc
     1801 ctccctaggc cctcctgtcc tgcaatgaca gaattaccgt tgtcgatggt cagcgaggag
     1861 gcagaagagc aagtccatcc cacccgaagc acatcttctg caaaccctgc tcggaggaaa
     1921 gcatgtattg tgaaggaaat ggaaaaaatg aagaacaagc gggaagagaa gagggcccag
     1981 aactcggaaa taagaataaa gcgagctcag gaatatgaca gcagctttcc aaactgggaa
     2041 tttgcccgga tgattaaaga attccgggtt actatagaat gtcacccact tactctgact
     2101 gatcctacgg aagagcacag gatctgcgtg tgtgtgagga agcgcccact gaataagcaa
     2161 gaactggcca agaaagaaat cgatgtgatt tctgttccca gcaagtgttt gctctttgtg
     2221 catgaaccca aattgaaagt ggacttgaca aagtatctgg agaatcaggc attctgcttc
     2281 gattttgcct ttgatgaaac agcttcaaat gaagtcgtct acaggttcac agcaaggcca
     2341 ctggtacaga cgatttttga agggggaaaa gcaacctgct ttgcctatgg gcagacgggc
     2401 agtgggaaga ctcacacaat gggtggagac ctgtctggta aatctcagaa tacatctaaa
     2461 gggatctatg caatggcttc ccgggatgtc ttccttctga agagtcagcc tcgctaccgg
     2521 aacttaaact tggaagttta tgtgacattc tttgagatct acaatgggaa ggtgtttgat
     2581 ctgctcaaca agaaggccaa gctacgtgta ctagaagaca gcaagcagca ggtgcaggtt
     2641 gtgggacttc aggagtacct ggttaactgt gctgatgatg tcatcaagat gctcaacatg
     2701 ggcagtgcct gcaggacttc tggacagact tttgccaact ccaattcttc ccgctcccat
     2761 gcctgcttcc agattcttct tcgagccaaa gggagattgc atggcaaatt ctccttggtg
     2821 gatctggcag ggaatgaacg aggggctgac acttctagtg ctgaccgcca gactcgcatg
     2881 gagggtgcag agattaacaa gtctctccta gcactgaagg agtgcatcag ggcgctggga
     2941 cagaacaagg ctgccacccc attcgctgag agcgcactga ctcaggtgct aagggattca
     3001 ttcattgggg agaactcgag gacttgcatg attgccatga tctcaccagg cataagctcc
     3061 tgtgaatata ctttaaacac gctgagatat gcagacagag tcaaggagct gagcccccac
     3121 agcgggctca gtggagagca gccaattcaa atggaaacgg aagagatgga agccagctct
     3181 aatgggacct cgctggcagt caatttcaag gaagaagagg agctgtcttc ccagatgtcc
     3241 agctttaatg aagccatgag tcagatcagg gagctggaag agagggccat ggaggagctc
     3301 agggagatca tacagcaagg accaggctgg cttgagctct ctgagatgac tgatcagcca
     3361 gactatgact tagagacctt tgtgaacaaa gcagaatctg ccctgactca gcagactaag
     3421 cacttctcag ccctgcgaga agtcatcaag gccctgcgtg tggccatgca actggaagaa
     3481 caggctagca aacagatgaa cagcaagaaa cggcaccaac accatcacca tcatcactaa
     3541 gcggccgcat cgataagctt cgaattctgc agtcgacggt accgcgggcc cgggatggcc
     3601 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3661 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3721 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3781 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     3841 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3901 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3961 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     4021 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     4081 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     4141 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     4201 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     4261 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     4321 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4381 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     4441 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4501 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4561 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4621 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4681 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4741 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     4801 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4861 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4921 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4981 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     5041 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5101 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5161 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5221 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5281 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5341 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5401 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5461 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5521 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5581 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5641 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5701 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5761 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     5821 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     5881 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5941 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     6001 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     6061 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6121 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6181 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6241 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6301 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6361 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6421 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6481 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6541 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6601 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6661 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6721 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6781 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6841 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6901 tgtggataac cgtattaccg ccatgcat
//
