LOCUS       sequence-10527-e        6420 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 6420)
  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 6420)
  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: pMX240
FEATURES             Location/Qualifiers
     source          1..6420
                     /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"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3208..3329
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3245..3264)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3299..3318
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3336..3791)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3473..3494)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3685..3704
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3818..3922
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3924..4281
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3924..4115
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3947..3967)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4132..4267
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4316..5110
                     /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             4316..5110
                     /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(4370..4389)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4980..4999
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5298..5317)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5342..5389
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5718..6306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6207..6226
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctatggg ccccaagcgg
     1381 cggcagctga cgttccggga gaagtcgcgg atcatccagg aggtggagga gaacccggac
     1441 ctacgcaagg gcgagatcgc gcggcgcttc aacatcccgc cgtccacgct gagcaccatc
     1501 ctgaagaaca agcgcgccat cctggcgtcg gagcgcaagt acggagtggc ctccacctgc
     1561 cgtaagacca acaagctgtc cccgtacgac aagctggagg ggcttctcat cgcttggttc
     1621 cagcagatcc gcgccgcggg cctgcctgtc aagggcatca tcctgaaaga gaaggcgcta
     1681 cggatagcgg aggagctggg catggacgac ttcacggctt ccaacggctg gctggatcgc
     1741 ttccgccggc gccacggtgt agtggcctgc agcggcgtga cccgctcccg ggcgcgaagc
     1801 tctaccccac gggccccagc ggcacctgcc ggcccagccg ccgtgccctc tgagggcagc
     1861 ggtggatccg ctgcaaaccc tgctcggagg aaagcatgta ttgtgaagga aatggaaaaa
     1921 atgaagaaca agcgggaaga gaagagggcc cagaactcgg aaataagaat aaagcgagct
     1981 caggaatatg acagcagctt tccaaactgg gaatttgccc ggatgattaa agaattccgg
     2041 gttactatag aatgtcaccc acttactctg actgatccta cggaagagca caggatctgc
     2101 gtgtgtgtga ggaagcgccc actgaataag caagaactgg ccaagaaaga aatcgatgtg
     2161 atttctgttc ccagcaagtg tttgctcttt gtgcatgaac ccaaattgaa agtggacttg
     2221 acaaagtatc tggagaatca ggcattctgc ttcgattttg cctttgatga aacagcttca
     2281 aatgaagtcg tctacaggtt cacagcaagg ccactggtac agacgatttt tgaaggggga
     2341 aaagcaacct gctttgccta tgggcagacg ggcagtggga agactcacac aatgggtgga
     2401 gacctgtctg gtaaatctca gaatacatct aaagggatct atgcaatggc ttcccgggat
     2461 gtcttccttc tgaagagtca gcctcgctac cggaacttaa acttggaagt ttatgtgaca
     2521 ttctttgaga tctacaatgg gaaggtgttt gatctgctca acaagaaggc caagctacgt
     2581 gtactagaag acagcaagca gcaggtgcag gttgtgggac ttcaggagta cctggttaac
     2641 tgtgctgatg atgtcatcaa gatgctcaac atgggcagtg cctgcaggac ttctggacag
     2701 acttttgcca actccaattc ttcccgctcc catgcctgct tccagattct tcttcgagcc
     2761 aaagggagat tgcatggcaa attctccttg gtggatctgg cagggaatga acgaggggct
     2821 gacacttcta gtgctgaccg ccagactcgc atggagggtg cagagattaa caagtctctc
     2881 ctagcactga aggagtgcat cagggcgctg ggacagaaca aggctcacac cccattccgt
     2941 gagagcaagc tgactcaggt gctaagggat tcattcattg gggagaactc gaggacttgc
     3001 atgattgcca tgatctcacc aggcataagc tcctgtgaat atactttaaa cacgctgaga
     3061 tatgcagaca gagtcaagga gctgagctaa tctagataac tgatcataat cagccatacc
     3121 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     3181 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     3241 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3301 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     3361 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     3421 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     3481 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     3541 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     3601 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3661 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3721 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     3781 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3841 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3901 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3961 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4021 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4081 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4141 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4201 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4261 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     4321 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4381 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4441 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     4501 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4561 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4621 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4681 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4741 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4801 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     4861 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4921 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4981 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5041 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5101 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     5161 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5221 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5281 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     5341 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     5401 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     5461 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     5521 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     5581 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5641 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5701 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5761 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5821 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     5881 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5941 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6001 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6061 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6121 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6181 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6241 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6301 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6361 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
