LOCUS       sequence-223978-        5199 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 5199)
  AUTHORS   Stumpff J, von Dassow G, Wagenbach M, Asbury C, Wordeman L
  TITLE     The kinesin-8 motor Kif18A suppresses kinetochore movements to 
            control mitotic chromosome alignment.
  JOURNAL   Dev Cell. 2008 Feb . 14(2):252-62.
  PUBMED    18267093
REFERENCE   2  (bases 1 to 5199)
  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: "Dev Cell. 
            2008 Feb . 14(2):252-62."
COMMENT     Sequence Label: pMX234
FEATURES             Location/Qualifiers
     source          1..5199
                     /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"
     polyA_signal    1987..2108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2024..2043)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2078..2097
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2115..2570)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2252..2273)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2464..2483
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2597..2701
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2703..3060
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2703..2894
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2726..2746)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2911..3046
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3095..3889
                     /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             3095..3889
                     /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(3149..3168)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3759..3778
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4077..4096)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4121..4168
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4497..5085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4986..5005
                     /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 atcctatggg ccccaagcgg cggcagctga cgttccggga gaagtcgcgg
     1381 atcatccagg aggtggagga gaacccggac ctacgcaagg gcgagatcgc gcggcgcttc
     1441 aacatcccgc cgtccacgct gagcaccatc ctgaagaaca agcgcgccat cctggcgtcg
     1501 gagcgcaagt acggagtggc ctccacctgc cgtaagacca acaagctgtc cccgtacgac
     1561 aagctggagg ggcttctcat cgcttggttc cagcagatcc gcgccgcggg cctgcctgtc
     1621 aagggcatca tcctgaaaga gaaggcgcta cggatagcgg aggagctggg catggacgac
     1681 ttcacggctt ccaacggctg gctggatcgc ttccgccggc gccacggtgt agtggcctgc
     1741 agcggcgtga cccgctcccg ggcgcgaagc tctaccccac gggccccagc ggcacctgcc
     1801 ggcccagccg ccgtgccctc tgagggcagc ggtggcagtt aagtcgacgg taccgcgggc
     1861 ccgggatggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt
     1921 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     1981 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2041 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2101 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     2161 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     2221 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     2281 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     2341 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     2401 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     2461 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     2521 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     2581 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2641 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2701 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     2761 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     2821 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2881 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     2941 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3001 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3061 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3121 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3181 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     3241 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     3301 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     3361 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     3421 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     3481 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     3541 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     3601 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     3661 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     3721 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     3781 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     3841 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     3901 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     3961 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4021 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     4081 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4141 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     4201 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     4261 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     4321 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     4381 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4621 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5161 tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
