LOCUS       sequence-171223-        5150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of stathmin-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5150)
  AUTHORS   Ringhoff DN, Cassimeris L
  TITLE     Stathmin regulates centrosomal nucleation of microtubules and 
            tubulin dimer/polymer partitioning.
  JOURNAL   Mol Biol Cell. 2009 Aug;20(15):3451-8. doi: 10.1091/mbc.E09-02-0140.
            Epub 2009 Jun 10.
  PUBMED    19515833
REFERENCE   2  (bases 1 to 5150)
  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"; doi: 
            "10.1091/mbc.E09-02-0140"; journalName: "Mol Biol Cell"; date: 
            "2009-08"; volume: "20"; issue: "15"; pages: "3451-8"
COMMENT     Sequence Label: stathmin-GFP
FEATURES             Location/Qualifiers
     source          1..5150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(42..61)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     389..410
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    578..699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(615..634)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     669..688
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(706..1161)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(843..864)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1055..1074
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1188..1292
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1294..1651
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1294..1485
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1317..1337)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1502..1637
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1686..2480
                     /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             1686..2480
                     /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(1740..1759)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2350..2369
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2668..2687)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2712..2759
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3088..3676
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3577..3596
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3851..4154
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4155..4358
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4309..4329
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4880..4889
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4886..4888,4889..4891,4892..5150,1..455)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4886..4888,4889..4891,4892..5150,1..452)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4931..4952)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
       61 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
      121 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
      181 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
      241 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
      301 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
      361 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
      421 ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct agatcataat
      481 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
      541 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
      601 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
      661 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
      721 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
      781 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
      841 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
      901 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
      961 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     1021 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     1081 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     1141 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1201 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1261 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     1321 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     1381 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     1441 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     1501 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     1561 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     1621 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     1681 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     1741 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     1801 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     1861 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     1921 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     1981 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     2041 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     2101 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     2161 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     2221 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     2281 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     2341 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     2401 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     2461 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     2521 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     2581 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     2641 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     2701 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     2761 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     2821 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     2881 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     2941 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3001 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3061 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3121 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3181 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3241 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3301 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3361 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3421 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3481 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3541 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3601 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3661 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3721 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3781 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     3841 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     3901 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     3961 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     4021 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     4081 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     4141 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     4201 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     4261 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     4321 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
     4381 gctagcgcta ccggactcag atctcgagct caagcttatg gcttcttctg atatccaggt
     4441 gaaagaactg gagaagcgtg cctcaggcca ggcttttgag ctgattctca gccctcggtc
     4501 aaaagaatct gttccagaat tccccctttc ccctccaaag aagaaggatc tttccctgga
     4561 ggaaattcag aagaaattag aagctgcaga agaaagacgc aagtcccatg aagctgaggt
     4621 cttgaagcag ctggctgaga aacgagagca cgagaaagaa gtgcttcaga aggcaataga
     4681 agagaacaac aacttcagta aaatggcaga agagaaactg acccacaaaa tggaagctaa
     4741 taaagagaac cgagaggcac aaatggctgc caaactggaa cgtttgcgag agaaggataa
     4801 gcacattgaa gaagtgcgga agaacaaaga atccaaagac cctgctgacg agactgaagc
     4861 tgacaaggat ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
     4921 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     4981 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     5041 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     5101 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
//
