LOCUS       sequence-456585-        6951 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 6951)
  AUTHORS   Ma N, Tulu US, Ferenz NP, Fagerstrom C, Wilde A, Wadsworth P
  TITLE     Poleward transport of TPX2 in the mammalian mitotic spindle requires
            dynein, Eg5, and microtubule flux.
  JOURNAL   Mol Biol Cell. 2010 Mar 15. 21(6):979-88.
  PUBMED    20110350
REFERENCE   2  (bases 1 to 6951)
  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. 2010 Mar 15. 21(6):979-88."
COMMENT     Sequence Label: pmCherry-TPX2
FEATURES             Location/Qualifiers
     source          1..6951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        53..356
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        357..560
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     511..531
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             605..1312
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(750..768)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1018..1037
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3731..3852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3768..3787)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3822..3841
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3859..4314)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3996..4017)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4208..4227
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4341..4445
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4447..4804
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4447..4638
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4470..4490)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4655..4790
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4839..5633
                     /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             4839..5633
                     /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(4893..4912)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5503..5522
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5821..5840)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5865..5912
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6241..6829
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6730..6749
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
       61 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      121 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      181 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      241 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      301 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      361 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      421 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      481 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      541 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggtcgc
      601 caccatggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
      661 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
      721 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
      781 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
      841 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
      901 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
      961 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     1021 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
     1081 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     1141 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     1201 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     1261 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtccggact
     1321 cagatctcga gctcaagctt cgaattctgc agtcgacatg tcacaagtta aaagctctta
     1381 ttcctatgat gccccctcgg atttcatcaa tttttcatcc ttggatgatg aaggagatac
     1441 tcaaaacata gattcatggt ttgaggagaa ggccaatttg gagaataagt tactggggaa
     1501 gaatggaact ggagggcttt ttcagggcaa aactcctttg agaaaggcta atcttcagca
     1561 agctattgtc acacctttga aaccagttga caacacttac tacaaagagg cagaaaaaga
     1621 aaatcttgtg gaacaatcca ttccgtcaaa tgcttgttct tccctggaag ttgaggcagc
     1681 catatcaaga aaaactccag cccagcctca gagaagatct cttaggcttt ctgctcagaa
     1741 ggatttggaa cagaaagaaa agcatcatgt aaaaatgaaa gccaagagat gtgccactcc
     1801 tgtaatcatc gatgaaattc taccctctaa gaaaatgaaa gtttctaaca acaaaaagaa
     1861 gccagaggaa gaaggcagtg ctcatcaaga tactgctgaa aagaatgcat cttccccaga
     1921 gaaagccaag ggtagacata ctgtgccttg tatgccacct gcaaagcaga agtttctaaa
     1981 aagtactgag gagcaagagc tggagaagag tatgaaaatg cagcaagagg tggtggagat
     2041 gcggaaaaag aatgaagaat tcaagaaact tgctctggct ggaatagggc aacctgtgaa
     2101 gaaatcagtg agccaggtca ccaaatcagt tgacttccac ttccgcacag atgagcgaat
     2161 caaacaacat cctaagaacc aggaggaata taaggaagtg aactttacat ctgaactacg
     2221 aaagcatcct tcatctcctg cccgagtgac taagggatgt accattgtta agcctttcaa
     2281 cctgtcccaa ggaaagaaaa gaacatttga tgaaacagtt tctacatatg tgccccttgc
     2341 acagcaagtt gaagacttcc ataaacgaac ccctaacaga tatcatttga ggagcaagaa
     2401 ggatgatatt aacctgttac cctccaaatc ttctgtgacc aagatttgca gagacccaca
     2461 gactcctgta ctgcaaacca aacaccgtgc acgggctgtg acctgcaaaa gtacagcaga
     2521 gctggaggct gaggagctcg agaaattgca acaatacaaa ttcaaagcac gtgaacttga
     2581 tcccagaata cttgaaggtg ggcccatctt gcccaagaaa ccacctgtga aaccacccac
     2641 cgagcctatt ggctttgatt tggaaattga gaaaagaatc caggagcgag aatcaaagaa
     2701 gaaaacagag gatgaacact ttgaatttca ttccagacct tgccctacta agattttgga
     2761 agatgttgtg ggtgttcctg aaaagaaggt acttccaatc accgtcccca agtcaccagc
     2821 ctttgcattg aagaacagaa ttcgaatgcc caccaaagaa gatgaggaag aggacgaacc
     2881 ggtagtgata aaagctcaac ctgtgccaca ttatggggtg ccttttaagc cccaaatccc
     2941 agaggcaaga actgtggaaa tatgcccttt ctcgtttgat tctcgagaca aagaacgtca
     3001 gttacagaag gagaagaaaa taaaagaact gcagaaaggg gaggtgccca agttcaaggc
     3061 acttcccttg cctcattttg acaccattaa cctgccagag aagaaggtaa agaatgtgac
     3121 ccagattgaa cctttctgct tggagactga cagaagaggt gctctgaagg cacagacttg
     3181 gaagcaccag ctggaagaag aactgagaca gcagaaagaa gcagcttgtt tcaaggctcg
     3241 tccaaacacc gtcatctctc aggagccctt tgttcccaag aaagagaaga aatcagttgc
     3301 tgagggcctt tctggttctc tagttcagga accttttcag ctggctactg agaagagagc
     3361 caaagagcgg caggagctgg agaagagaat ggctgaggta gaagcccaga aagcccagca
     3421 gttggaggag gccagactac aggaggaaga gcagaaaaaa gaggagctgg ccaggctacg
     3481 gagagaactg gtgcataagg caaatccaat acgcaagtac cagggtctgg agataaagtc
     3541 aagtgaccag cctctgactg tgcctgtatc tcccaaattc tccactcgat tccactgcta
     3601 aggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     3661 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3721 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3781 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3841 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3901 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3961 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4021 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4081 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4141 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4201 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4261 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     4321 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4381 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4441 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4501 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4561 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4621 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4681 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4741 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4801 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4861 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4921 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4981 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     5041 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5101 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5161 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5221 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5281 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5341 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     5401 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5461 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5521 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5581 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5641 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5701 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5761 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     5821 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5881 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5941 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     6001 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     6061 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     6121 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6181 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6241 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6301 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6361 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6421 caagaactct gtatcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6481 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6541 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6601 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6661 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6721 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6781 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6841 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6901 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat t
//
