LOCUS       sequence-328015-        8540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged KIF5A protein in pcDNA3.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8540)
  AUTHORS   Fukuda Y, Pazyra-Murphy MF, Silagi ES, Tasdemir-Yilmaz OE, Li Y, 
            Rose L, Yeoh ZC, Vangos NE, Geffken EA, Seo HS, Adelmant G, Bird GH,
            Walensky LD, Marto JA, Dhe-Paganon S, Segal RA
  TITLE     Binding and transport of SFPQ-RNA granules by KIF5A/KLC1 motors 
            promotes axon survival.
  JOURNAL   J Cell Biol. 2021 Jan 4;220(1). pii: 211577. doi: 
            10.1083/jcb.202005051.
  PUBMED    33284322
REFERENCE   2  (bases 1 to 8540)
  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 
            Biol."; date: "2021-01-4"; volume: "220(1). pii: 211577. doi"; 
            pages: " 10.1083/jcb.202005051"
COMMENT     Sequence Label: HA-KIF5A
FEATURES             Location/Qualifiers
     source          1..8540
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             708..734
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3897..3914)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3903..4127
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4173..4601
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4260..4279)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4470..4491
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4610..4630)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4615..4944
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4795..4930
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4857..4876
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5011..5805
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5065..5084)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5675..5694
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5979..6100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6016..6035)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6070..6089
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6149..6165)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6149..6165)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6162..6184)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6173..6189
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6197..6203,6204..6221,6222..6227))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6242..6263
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6380..6397)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6551..7139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6631..6650)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7310..8101,8102..8170))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7933..7952
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8171..8275)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8350..8369)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc caccatgtac ccatacgatg
      721 ttccagatta cgctggatcc ggaatggcgg agactaacaa cgaatgcagc atcaaggtgc
      781 tttgccgatt tcggcccctg aaccaggccg agattctgcg gggggacaag ttcatcccca
      841 ttttccaagg ggacgacagc gtcattattg ggggaaagcc atatgtcttt gaccgcgtct
      901 tccccccaaa caccactcag gagcaggttt accacgcctg tgccatgcag atcgtcaaag
      961 acgtccttgc tggttacaat ggcacaatct tcgcttatgg acagacatcc tcagggaaaa
     1021 cgcataccat ggaggggaag ctgcacgacc ctcagctgat gggcatcatt ccccggatcg
     1081 ctcgagacat cttcaaccac atctactcca tggatgagaa ccttgaattc cacattaagg
     1141 tatcttactt cgagatttac ctggataaga tccgtgacct tttggatgtg accaagacga
     1201 acctgtccgt gcatgaggac aaaaaccggg tgccgtttgt caagggttgt accgaacgct
     1261 ttgtgtccag cccagaggag attctggatg tgatcgatga ggggaagtcc aaccgtcacg
     1321 tagctgtcac caacatgaac gagcacagtt ctcggagcca cagcatcttc ctcatcaaca
     1381 tcaagcagga gaacgtagag accgagcaga agctcagcgg gaagctgtac ctcgtggatc
     1441 tggccggaag cgagaaggtc agcaagacag gggcagaggg agccgttctg gacgaggcaa
     1501 agaatatcaa caagtcgctg tcggccctgg ggaacgtgat ctctgcactg gcagagggca
     1561 ccaaaagcta tgtgccgtac cgcgacagca aaatgacgag gattctccag gactctctgg
     1621 aagggaactg caggactacc atgttcatct gctgctcgcc gtccagctac aatgacgcag
     1681 agaccaagtc cacgctcatg tttgggcagc gggcgaagac catcaagaac actgcctcag
     1741 tgaatctgga gctgactgct gagcagtgga agaagaagta tgagaaggag aaggagaaga
     1801 ccaagaccca gaaggagaca attgcgaagc tagaggctga gcttagccgg tggcgcaatg
     1861 gagagaatgt gcctgagact gagcgcctgg ctggagagga ctcagctctg ggagctgagc
     1921 tctgcgagga gacccctgtg aatgacaact catccattgt ggtacgcatc gcacctgagg
     1981 aaaggcagaa atatgaggaa gagatccgcc gtctctacaa gcagcttgat gacaaggatg
     2041 atgagatcaa ccagcagagc cagctcattg agaagctgaa gcagcagatg ctggaccagg
     2101 aagagctgct cgtgtccact cggggagaca acgagaaggt ccagcgggag cttagccacc
     2161 tgcagtccga gaacgatgct gcgaaggacg aggtgaagga agtgctgcag gccctagagg
     2221 agctggcggt caactacgac cagaagtccc aggaggtgga ggagaagagc cagcagaacc
     2281 agctgctggt ggacgagctg tcccagaaag tggccaccat gctgtccctg gagtccgagc
     2341 tacagcggct ccaggaggtc agtggacacc agcgaaagcg gatcgctgag gtgctgaatg
     2401 ggctgatgag ggacctgagt gagttcagtg tcatcgtggg caacggcgag attaagctgc
     2461 ccgtggagat cagtggggcc atcgaggagg agttcacggt ggcccggctc tacatcagca
     2521 agatcaagtc ggaggtgaag tccgtggtta agcgatgtcg gcagctggag aacctccagg
     2581 tggagtgtca tcgcaagatg gaggtgaccg gtagggagct gtcatcttgc caactgctca
     2641 tctcacagca tgaggccaag atccgttcac tcacggagta catgcagact gtggagttga
     2701 agaaacggca cctggaagag tcctacgact ccctgagcga tgagcttgcc aggctccagg
     2761 cgcacgaaac tgtacacgag gtagctctga aagacaagga gccagacaca caggacgcgg
     2821 aggaggtgaa gaaggccctg gaactacaga tggagaatca ccgtgaggcc catcaccggc
     2881 agctggcccg cctccgggat gagattaatg agaaacagaa aaccattgat gagctgaaag
     2941 acctgaacca gaagctccag ttagagctgg agaagcttca ggccgactat gagaggctga
     3001 agaatgaaga gaatgagaag agcgccaagc tccaggagct gacatttctg tatgagcgac
     3061 atgagcagtc caagcaggac ctcaaggggc tggaggagac agttgcccgt gaactccaga
     3121 ccctccacaa ccttcgcaag ctgttcgttc aagacgtcac gactcgagtc aagaaaagtg
     3181 cagaaatgga gcccgaggac agtgggggga ttcattccca aaagcagaag atctcctttc
     3241 ttgagaacaa cctggaacag cttacaaagg ttcacaaaca gctggtacgt gacaatgcag
     3301 atctgcgttg tgagcttcct aaattggaaa aacgacttcg ggctacggct gagagagtta
     3361 aggccctgga gggtgcactg aaggaggcca aggagggtgc tatgaaggac aagcgtagat
     3421 accagcagga ggtggaccgc atcaaggaag ccgtgcggta caagagctcc ggcaagcggg
     3481 gccattctgc ccagatcgct aagcctgtga ggcctggcca ctatcctgcc tcctcaccca
     3541 ccaaccccta cggcacccgg agccccgagt gtatcagcta caccaacaac ctcctccaga
     3601 actaccagaa cctgcacctg caggctgcgc ctagctccac ttcagatatg tactttgcca
     3661 gtagcggagc cacatctgtt gcccccttgg cttcctacca gaaggccaac atggacaatg
     3721 gaaatgccac agatatcaac gacaacagga gtgacctgcc gtgtggctat gaggctgagg
     3781 accaggccaa gcttttccct ctccaccaag agacagcagc cagttagaat tctgcagata
     3841 tccagcacag tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc
     3901 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3961 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     4021 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     4081 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     4141 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     4201 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4261 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4321 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4381 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4441 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4501 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4561 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     4621 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4681 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4741 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4801 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4861 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4921 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4981 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     5041 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     5101 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     5161 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     5221 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5281 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5341 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5401 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5461 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5521 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5581 gcctgcttgc craatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5641 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5701 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5761 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5821 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5881 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5941 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     6001 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6061 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     6121 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     6181 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     6241 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6301 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6361 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6421 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6481 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6541 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6601 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6661 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6721 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6781 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6841 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6901 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6961 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     7021 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7081 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7141 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7201 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7261 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7321 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7381 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7441 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7501 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7561 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7621 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7681 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7741 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7801 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7861 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7921 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7981 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8041 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8101 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8161 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8221 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8281 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg
     8341 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     8401 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     8461 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
     8521 acgggccaga tatacgcgtt
//
