LOCUS       sequence-13751-e        6759 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 6759)
  AUTHORS   Manning AL, Ganem NJ, Bakhoum SF, Wagenbach M, Wordeman L, Compton 
            DA
  TITLE     The kinesin-13 proteins Kif2a, Kif2b, and Kif2c/MCAK have distinct 
            roles during mitosis in human cells.
  JOURNAL   Mol Biol Cell. 2007 Aug . 18(8):2970-9.
  PUBMED    17538014
REFERENCE   2  (bases 1 to 6759)
  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: "Mol Biol 
            Cell. 2007 Aug . 18(8):2970-9."
COMMENT     Sequence Label: pMX230
FEATURES             Location/Qualifiers
     source          1..6759
                     /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"
     CDS             3388..3405
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3547..3668
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3584..3603)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3638..3657
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3675..4130)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3812..3833)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4024..4043
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4157..4261
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4263..4620
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4263..4454
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4286..4306)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4471..4606
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4655..5449
                     /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             4655..5449
                     /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(4709..4728)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5319..5338
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5637..5656)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5681..5728
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6057..6645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6546..6565
                     /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 attccaccat ggccagccag
     1381 ttctgcctcc ctgaatcccc atgtctctcg cccctgaaac ccttgaagcc acatttcgga
     1441 gacatccaag agggcatcta cgtggcgatc cagcgcagtg acaagcggat ccacctcgct
     1501 gtggtcacgg agatcaacag agaaaactat tgggtcacgg tagagtgggt ggagaaagca
     1561 gtcaaaaaag gcaagaagat tgacctggag accatactcc tgctgaatcc agctctggac
     1621 tctgctgaac accccatgcc gcccccgccc ttatccccct tggctctggc gccctcttcg
     1681 gccatcaggg accagcgtac cgccacgaaa tgggttgcga tgatccccca gaaaaaccaa
     1741 acagcctcag gggacagcct ggatgtgagg gtccccagca aaccttgtct gatgaagcag
     1801 aaaaagtctc cctgcctctg ggaaatccag aaactgcagg agcagcggga aaagcgcagg
     1861 cggctgcagc aggagatccg agctagacgc gccctcgatg tcaataccag aaaccccaac
     1921 tacgaaatca tgcacatgat cgaagagtat cgcaggcacc tggacagcag caagatctca
     1981 gtcctggagc ccccgcaaga acatcgcatc tgcgtctgcg tgaggaagcg gcctctcaac
     2041 cagcgagaga caaccttgaa ggacctggat atcatcaccg tcccctcgga caatgtggtt
     2101 atggtgcatg agtccaagca aaaggtggac ctcactcgct acctgcagaa ccagaccttc
     2161 tgcttcgacc atgccttcga tgacaaagcc tccaacgagt tggtgtacca gttcaccgcc
     2221 cagccactgg tggagtccat cttccgcaag ggcatggcca cctgctttgc ctatgggcag
     2281 acgggaagtg ggaagacgta caccatgggt ggagactttt caggaacggc ccaagattgt
     2341 tctaagggca tttatgcttt ggtggcacag gatgtctttc tcctgctcag aaactccaca
     2401 tatgagaagc tggacctcaa agtctatggg acattttttg agatttatgg gggcaaggtg
     2461 tatgatttgt tgaactggaa gaagaagctg caagtccttg aggatggcaa tcagcaaatc
     2521 caagtggtcg ggctgcagga gaaagaggtg tgttgtgtgg aggaagtgct gaacctggtg
     2581 gaaataggga atagctgtcg gacttccagg caaacatctg tcaacgctca ctcatccagg
     2641 agccatgcag tgttccagat catcctgaag tcaggaggga taatgcatgg caagttttcc
     2701 ctcgttgatt tagctgggaa tgaaagagga gcagatacaa ccaaggccag ccggaaaagg
     2761 cagctggaag gggcagagat taacaagagt cttctagccc tcaaagaatg tattctggct
     2821 ttgggtcaga acaagcctca caccccattc agagccagca aactcacact ggtgctccgg
     2881 gactccttta taggccagaa ctcctccact tgcatgattg ctaccatctc tccggggatg
     2941 acctcttgtg aaaacactct caacacttta agatatgcaa acagagtaaa aaaattaaat
     3001 gtagatgtaa ggccctacca tcgtggccac tatccgattg gacatgaggc accaaggatg
     3061 ttaaaaagtc acatcggaaa ttcagaaatg tcccttcaga gggatgaatt tattaaaata
     3121 ccttatgtac agagtgagga gcagaaagag attgaagagg ttgaaacatt acccactctg
     3181 ttagggaagg ataccacaat ttcagggaag ggatctagcc aatggttgga aaacatccag
     3241 gagagagctg gtggagtaca ccatgatatt gatttttgca ttgcccggtc tttgtccatt
     3301 ttggagcaga aaattgatgc tctgaccgag atccaaaaga aactgaaatt attactagct
     3361 gacctccacg tgaagagcaa ggtagagcat caccaccatc accattaagc ggccgcggga
     3421 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     3481 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3541 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3601 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3661 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3721 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3781 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3841 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3901 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3961 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     4021 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     4081 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     4141 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4201 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4261 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4321 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4381 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4441 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4501 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4561 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4621 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4681 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4741 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4801 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4861 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4921 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4981 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5041 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5101 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5161 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5221 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5281 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5341 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5401 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5461 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5521 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5581 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     5641 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5701 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5761 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5821 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     5881 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     5941 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6001 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6061 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6121 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6181 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     6241 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6301 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6361 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6421 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6481 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6541 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6601 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6661 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6721 tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
