LOCUS       sequence-171227-        5150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of stathmin-CFP.
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-CFP
FEATURES             Location/Qualifiers
     source          1..5150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        47..404
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        47..238
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(70..90)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      255..390
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             439..1233
                     /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             439..1233
                     /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(493..512)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1103..1122
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1421..1440)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1465..1512
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1841..2429
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2330..2349
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2604..2907
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2908..3111
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3062..3082
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3633..3642
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3639..3641,3642..3644,3645..4358)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3684..3705)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3945..3964)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4292..4313
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4481..4602
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4518..4537)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4572..4591
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4609..5064)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4746..4767)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4958..4977
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5091..5150,1..45)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
       61 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
      121 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
      181 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
      241 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
      301 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
      361 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
      421 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
      481 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
      541 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
      601 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
      661 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
      721 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
      781 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
      841 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
      901 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
      961 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     1021 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     1081 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     1141 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     1201 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     1261 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     1321 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     1381 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     1441 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     1501 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     1561 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     1621 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     1681 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1741 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1801 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1861 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1921 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1981 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2041 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2101 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2161 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2221 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2281 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2341 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2401 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2461 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2521 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     2581 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     2641 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     2701 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     2761 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2821 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     2881 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     2941 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     3001 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     3061 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     3121 aaccgtcaga tccgctagcg ctaccggact cagatctcga gctcaagctt atggcttctt
     3181 ctgatatcca ggtgaaagaa ctggagaagc gtgcctcagg ccaggctttt gagctgattc
     3241 tcagccctcg gtcaaaagaa tctgttccag aattccccct ttcccctcca aagaagaagg
     3301 atctttccct ggaggaaatt cagaagaaat tagaagctgc agaagaaaga cgcaagtccc
     3361 atgaagctga ggtcttgaag cagctggctg agaaacgaga gcacgagaaa gaagtgcttc
     3421 agaaggcaat agaagagaac aacaacttca gtaaaatggc agaagagaaa ctgacccaca
     3481 aaatggaagc taataaagag aaccgagagg cacaaatggc tgccaaactg gaacgtttgc
     3541 gagagaagga taagcacatt gaagaagtgc ggaagaacaa agaatccaaa gaccctgctg
     3601 acgagactga agctgacaag gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc
     3661 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     3721 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     3781 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctggg
     3841 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     3901 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     3961 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     4021 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacatca
     4081 gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga
     4141 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     4201 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     4261 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     4321 ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat
     4381 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     4441 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     4501 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4561 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     4621 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     4681 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     4741 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     4801 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     4861 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     4921 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     4981 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     5041 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     5101 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
//
