LOCUS       sequence-175297-        6166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of rat GFP/Katanin A subunit in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6166)
  AUTHORS   Karabay A, Yu W, Solowska JM, Baird DH, Baas PW
  TITLE     Axonal growth is sensitive to the levels of katanin, a protein that 
            severs microtubules.
  JOURNAL   J Neurosci. 2004 Jun 23;24(25):5778-88.
  PUBMED    15215300
REFERENCE   2  (bases 1 to 6166)
  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 
            Neurosci."; date: "2004-06-23"; volume: "24(25)"; pages: "5778-88"
COMMENT     Sequence Label: GFP/KATNA1
FEATURES             Location/Qualifiers
     source          1..6166
                     /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"
     polyA_signal    2954..3075
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2991..3010)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3045..3064
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3082..3537)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3219..3240)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3431..3450
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3564..3668
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3670..4027
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3670..3861
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3693..3713)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3878..4013
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4062..4856
                     /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             4062..4856
                     /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(4116..4135)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4726..4745
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5044..5063)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5088..5135
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5464..6052
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5953..5972
                     /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 atccagtctt ctaatgatta ctgagaatgt aaaattggct
     1381 cgtgagtatg cattgctggg aaactatgac tctgcaatgg tctattatca gggagttctt
     1441 gaccaaatta acaagtatct atactcagtc aaagatacac acctccatca gaaatggcaa
     1501 caggtttggc aggaaataaa cgtggaagct aagcatgtta aggagatcat gaaaacactg
     1561 gagagcttta aactggacag cacttctttg aaagctgcac agcatgagct tccgtcctcg
     1621 gaaggagaag tctggtcttt gccggtacct gttgaaagga gacccctacc aggacctagg
     1681 aagcgccagt ctactcagca cagtgaccct aagccacaca gtaaccggcc aggcgcagtc
     1741 gtcagagctc atcgaccatc tgcacagagt ctgcacagcg acagaggcaa agctgttcgt
     1801 agtcgtgaaa agaaagagca gagtaagggc agagaggaaa agaacaaatt acctgctgca
     1861 gttacagaac cagaggcaaa taaatttgat agcacggggt atgacaagga cttagtagaa
     1921 gctttggaaa gagatataat ttctcagaat cccaacgttc ggtggtatga tattgctgat
     1981 ttagtagaag ctaaaaagtt gcttcaggaa gcagtggtgt taccaatgtg gatgccagaa
     2041 ttctttaagg gcattaggag accatggaaa ggtgtgctga tggttggccc acctggcact
     2101 ggaaagaccc tccttgctaa agcagttgcc acagaatgca agacaacgtt cttcaatgtc
     2161 tcttcatcca ctctgacttc caaatacaga ggagagtctg agaagcttgt ccgtcttctg
     2221 tttgaaatgg ctcggtttta ctccccagct accatattta ttgatgagat agattccatc
     2281 tgcagccgcc gaggaacttc tgaagagcat gaagcaagcc gaagggtgaa agctgagctg
     2341 ctggttcaga tggatggtgt tggaggtgcc tcagaaaatg acgacccttc caagatggtc
     2401 atggttctgg cggccactaa tttcccatgg gacatagatg aggctttaag gcgtcgtctg
     2461 gagaaaagaa tctacatacc gttaccatca gcaaaaggca gagaagaact gttaagaata
     2521 agtctacgag agctggaatt ggctgatgat gttaaccttg caagcatagc agaaaacatg
     2581 gaaggctact caggtgcaga cattaccaat gtatgcaggg atgcgtcctt gatggccatg
     2641 agacgccgta ttgagggtct aaccccagaa gaaatccgga acctttcaag agaagaaatg
     2701 cacatgccta ccaccatgga ggattttgaa atggctttaa agaaggtctc caaatcagta
     2761 tcagctgcag acattgagag atacgagaaa tggatagttg agtttggatc atgctaaggg
     2821 cccgggatcc accggatcta gataactgat cataatcagc cataccacat ttgtagaggt
     2881 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     2941 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3001 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3061 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     3121 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     3181 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     3241 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     3301 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     3361 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3421 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3481 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     3541 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3601 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3661 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     3721 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3781 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3841 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3901 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3961 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4021 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4081 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4141 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4201 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     4261 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4321 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4381 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4441 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4501 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4561 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     4621 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4681 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4741 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4801 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4861 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4921 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4981 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     5041 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     5101 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     5161 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     5221 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     5281 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     5341 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5401 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5461 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5521 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5581 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5641 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5701 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5761 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5821 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5881 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5941 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6001 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6061 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6121 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
