LOCUS       sequence-10529-e        6010 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 6010)
  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 6010)
  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: pMX232
FEATURES             Location/Qualifiers
     source          1..6010
                     /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             2593..2610
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2798..2919
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2835..2854)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2889..2908
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2926..3381)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3063..3084)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3275..3294
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3408..3512
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3514..3871
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3514..3705
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3537..3557)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3722..3857
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3906..4700
                     /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             3906..4700
                     /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(3960..3979)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4570..4589
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4888..4907)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4932..4979
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5308..5896
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5797..5816
                     /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 aacgcagatc agtcaactcc aaaattccag ctccaaaaga aggtcttcga
     1681 agccgttcca ctcgcatgtc cactgtccca 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 tcatgtattg 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 cgtgagcccc cacagcgggc tcagtggaga gcagccaatt
     2221 caaatggaaa cggaagagat ggaagccagc tctaatggga cctcgctggc agtcaatttc
     2281 aaggaagaag aggagctgtc ttcccagatg tccagcttta atgaagccat gagtcagatc
     2341 agggagctgg aagagagggc catggaggag ctcagggaga tcatacagca aggaccaggc
     2401 tggcttgagc tctctgagat gactgatcag ccagactatg acttagagac ctttgtgaac
     2461 aaagcagaat ctgccctgac tcagcagact aagcacttct cagccctgcg agaagtcatc
     2521 aaggccctgc gtgtggccat gcaactggaa gaacaggcta gcaaacagat gaacagcaag
     2581 aaacggcacc aacaccatca ccatcatcac taagcggccg catcgataag cttcgaattc
     2641 tgcagtcgac ggtaccgcgg gcccgggatc gacgcgtcgg ccgcgactct agatcataat
     2701 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     2761 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     2821 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2881 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     2941 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3001 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3061 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3121 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3181 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3241 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3301 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3361 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3421 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3481 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     3541 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3601 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3661 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3721 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3781 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3841 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     3901 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3961 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4021 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4081 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4141 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4201 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4261 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4321 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4381 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     4441 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4501 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4561 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4621 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4681 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4741 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4801 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4861 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     4921 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     4981 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     5041 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     5101 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     5161 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5221 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5281 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5341 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5401 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     5461 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5521 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5581 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5641 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5701 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5761 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5821 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5881 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5941 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6001 cgccatgcat
//
