LOCUS       sequence-322835-        7746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of GFP-Kif5c.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7746)
  AUTHORS   Dunn S, Morrison EE, Liverpool TB, Molina-Paris C, Cross RA, Alonso 
            MC, Peckham M
  TITLE     Differential trafficking of Kif5c on tyrosinated and detyrosinated 
            microtubules in live cells.
  JOURNAL   J Cell Sci. 2008 Apr 1;121(Pt 7):1085-95. doi: 10.1242/jcs.026492. 
            Epub 2008 Mar 11.
  PUBMED    18334549
REFERENCE   2  (bases 1 to 7746)
  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: "J Cell 
            Sci."; date: "2008-04-1"; volume: "121(Pt 7):1085-95. doi"; pages: "
            10.1242/jcs.026492. Epub 2008 Mar 11"
COMMENT     Sequence Label: pGFP-Kif5c
FEATURES             Location/Qualifiers
     source          1..7746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1310..1319
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1316..1318,1319..1321,1322..2032)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1316..1318,1319..1321,1322..2032)
                     /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(1361..1382)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1622..1641)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1969..1990
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2038..2094
                     /label=MCS
                     /note="multiple cloning site"
     CDS             2962..2973
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    5237..5358
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5274..5293)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5328..5347
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5365..5820)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5502..5523)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5714..5733
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5847..5951
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5953..6310
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5953..6144
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5976..5996)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6161..6296
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6345..7139
                     /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             6345..7139
                     /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(6399..6418)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7009..7028
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7327..7346)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7371..7418
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt
     1321 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1381 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1441 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1501 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     1561 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1621 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1681 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1741 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     1801 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     1861 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1921 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1981 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtactcaga
     2041 tctcgagctc aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccagccga
     2101 atgcagcatc aaagtgatgt gccggttccg gcccctcaac gaagctgaga tcctccgcgg
     2161 ggacaaattc atccccaaat tcaaaggcga ggagacggtg gtgatcgggc aagggaagcc
     2221 gtatgtcttt gaccgagttc tgccgcccaa cacaacccag gagcaggtct acaatgcctg
     2281 tgcgaagcag attgtcaaag atgtcctcga gggttacaat gggacaattt ttgcatacgg
     2341 gcagacttca tcaggaaaaa ctcataccat ggaggggaag ctgcacgatc ctcagcttat
     2401 gggtatcata cccaggatcg cacacgatat tttcgatcac atctattcca tggatgagaa
     2461 cttggagttt catatcaagg tttcctattt tgagatctac ttggacaaaa taagggactt
     2521 gcttgacgtg tccaagacca acttggctgt tcatgaagac aaaaacagag tcccctatgt
     2581 aaaggggtgc accgagaggt ttgtgtcaag cccggaggag gtcatggatg tgatcgatga
     2641 gggcaaagca aaccgacatg tggctgtgac aaacatgaac gaacacagct ccaggagtca
     2701 cagtatcttc ctgattaaca ttaaacaaga gaatgtggag actgaaaaaa aactcagcgg
     2761 gaagctgtat ttggttgatt tggctgggag tgaaaaggtc agcaaaactg gtgccgaggg
     2821 agctgttctt gatgaagcta aaaatatcaa caagtctttg tctgctcttg gaaatgtgat
     2881 ctctgccttg gcagaaggga caaaaacaca tgtgccatac cgggacagca agatgactcg
     2941 gattctccag gactctctgg gtgggaactg caggaccacc attgtcattt gctgttctcc
     3001 ttcagtcttc aatgaggctg agaccaagtc cacgctgatg tttggacaga gagcaaagac
     3061 catcaagaat acagtctctg tgaacttgga actaacagca gaagaatgga agaagaaata
     3121 tgaaaaagag aaagagaaga ataaggccct gaagagtgtc atccagcatc tggaggtgga
     3181 gctgaacagg tggaggaacg gggaagctgt gcctgaggat gaacagatca gtgccaagga
     3241 ccagaagaac ctggagccct gcgacaacac acccatcata gacaacatca cacctgtggt
     3301 tgacggcatc tctgctgaga aagagaagta tgacgaggaa atcaccagtc tgtaccgaca
     3361 gctggatgac aaggacgatg aaattaacca gcagagccag ctggctgaaa agctgaagca
     3421 acagatgttg gatcaggacg agcttctagc ttccaccaga agggactacg agaaaatcca
     3481 ggaggagctg acccgcctcc agattgaaaa cgaggcggct aaagacgagg tgaaggaggt
     3541 cctccaggcc ctggaggagc tggctgtcaa ttacgaccag aagtcacagg aagtggagga
     3601 caagacccgg gccaacgagc aactgaccga tgagctggcc cagaaaacga cgacactgac
     3661 aaccacccag cgggagctga gtcagctaca ggagcttagc aaccaccaga agaagagggc
     3721 cacggaaatc ctgaacctgc ttctcaagga tctgggcgag ataggtggaa ttatcggcac
     3781 caacgatgtg aagaccctgg cagatgtgaa tggagtcatc gaagaggagt tcaccatggc
     3841 ccgcctgtac attagcaaga tgaagtcgga ggtcaagtct ctcgtgaacc gcagcaagca
     3901 gctggagagc gcccagacgg actccaacag gaagatgaat gccagcgagc gggagctggc
     3961 agcttgccag ttgcttatct cgcagcatga agccaagatc aagtctctga cagactacat
     4021 gcagaacatg gaacagaaga ggcggcagct ggaagagtcg caggactccc tcagcgagga
     4081 gctggccaag ctccgggccc aagaaaaaat gcacgaagtc agtttccaag ataaggaaaa
     4141 ggagcacctg acgaggctgc aggatgctga ggaagtgaag aaggctctgg agcagcagat
     4201 ggagagtcac cgggaagcgc accaaaagca gctgtccaga cttcgtgatg agattgagga
     4261 gaagcagaga atcattgatg agattcggga tttgaatcag aagctgcaac tggaacagga
     4321 gaggctcagc tctgattata acaagctgaa aatagaggac caggagagag aagtgaagct
     4381 ggagaagctc ctattgctca atgacaaaag agagcaagcc agggaggacc tcaagggact
     4441 ggaagagact gtgtccagag aactccagac tcttcacaac ctgcgcaaac tcttcgtcca
     4501 ggacttgacc acccgggtga aaaagagtgt ggagctggac agcgacgatg gagggggtag
     4561 tgccgcccag aagcagaaga tctccttcct ggagaacaac ctggagcagc ttaccaaggt
     4621 gcacaagcag ctggtccggg acaatgcgga tttgcgctgt gaactcccca agctggagaa
     4681 gaggctccgt gctactgcag agcgcgtcaa ggccttggag agtgcgctca aagaggccaa
     4741 ggagaatgcc atgagggacc gaaagcgcta ccagcaggaa gtggatcgca tcaaggaggc
     4801 tgtgcgggcc aagaacatgg ccaggagggc acattcggct cagatcgcca agcccatccg
     4861 cccaggacac tatcctgcat catctccaac agccgtccat gccgtccggg gaggaggtgg
     4921 atcttcaaac tccactcact accagaagta aatgcagagt gagtccctgc agcctggggt
     4981 gttttctctt cacagcaacc ccagtgctta tgaggcatag ccctcttgcc gctctgaaac
     5041 cccaccctgg ccttcagccc tattatctaa tagagattag ttcagctctg ttccaggaag
     5101 aggcctggga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
     5161 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     5221 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5281 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5341 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     5401 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     5461 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     5521 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     5581 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     5641 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     5701 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     5761 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     5821 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5881 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5941 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     6001 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     6061 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6121 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     6181 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     6241 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     6301 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     6361 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     6421 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     6481 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     6541 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     6601 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     6661 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     6721 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     6781 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     6841 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     6901 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     6961 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     7021 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     7081 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     7141 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     7201 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     7261 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     7321 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     7381 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     7441 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     7501 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     7561 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     7621 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7681 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7741 atcttc
//
