LOCUS       sequence-444944-        7639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of CLIPf-KIF5A(1-573).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7639)
  AUTHORS   Smith ER, Turner ED, Abdelhamid MAS, Craggs TD, Twelvetrees AE
  TITLE     Kinesin-1 is highly flexible and adopts an open conformation in the 
            absence of cargo
  JOURNAL   bioRxiv 2024.12.20.629623
REFERENCE   2  (bases 1 to 7639)
  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: "bioRxiv 
            2024.12.20.629623"
COMMENT     Sequence Label: CLIPf-KIF5A(1-573)
FEATURES             Location/Qualifiers
     source          1..7639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     34..53
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(272..376)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(451..470)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        642..1021
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1022..1225
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1176..1196
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1270..1289
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1270..1288
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3637..3654)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3643..3867
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3913..4341
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4000..4019)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4210..4231
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4350..4370)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4355..4684
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4535..4670
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4597..4616
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4751..5545
                     /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
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLCEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4805..4824)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5415..5434
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5719..5840
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5756..5775)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5810..5829
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5889..5905)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5889..5905)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5902..5924)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5913..5929
                     /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(5937..5943,5944..5961,5962..5967))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5982..6003
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6120..6137)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6291..6879)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6371..6390)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7050..7639,1..202,203..271))
                     /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"
ORIGIN
        1 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
       61 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
      121 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
      181 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
      241 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
      301 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
      361 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
      421 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
      481 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
      541 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
      601 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
      661 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      721 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      781 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      841 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      901 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      961 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1021 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1081 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1141 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1201 gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg cttactggct
     1261 tatcgaaatt aatacgactc actataggga gacccaagct ggctagcgtt taaacttaag
     1321 cttggtacca tggacaagga ctgcgagatg aagcggacca cactggatag ccctctgggc
     1381 aagctggaac tgagcggctg tgaacagggc ctgcaccgga tcatctttct cggcaaggga
     1441 acctctgccg ccgatgctgt tgaagtgcct gctcctgctg ctgtgctcgg aggacctgaa
     1501 cctctgattc aggccaccgc ctggctgaac gcctactttc atcagcccga ggccatcgag
     1561 gagttccccg ttcctgctct gcatcacccc gtgttccagc aagaaagctt caccagacag
     1621 gtgctgtgga agctgctgaa ggtggtcaag ttcggcgaag tgatcagcga gtctcacctg
     1681 gccgctctcg tgggaaatcc agctgctaca gccgccgtga acacagccct ggatggaaac
     1741 cctgtgccta ttctgatccc ctgccacaga gtggtgcagg gcgattctga tgtgggccct
     1801 tatcttggag gcctggccgt gaaagaatgg ctgctggccc acgaaggaca cagactggga
     1861 aaacctggcc tcggaggatc catggcggag actaacaacg aatgcagcat caaggtgctt
     1921 tgccgatttc ggcccctgaa ccaggccgag attctgcggg gggacaagtt catccccatt
     1981 ttccaagggg acgacagcgt cattattggg ggaaagccat atgtctttga ccgcgtcttc
     2041 cccccaaaca ccactcagga gcaggtttac cacgcctgtg ccatgcagat cgtcaaagac
     2101 gtccttgctg gttacaatgg cacaatcttc gcttatggac agacatcctc agggaaaacg
     2161 cataccatgg aggggaagct gcacgaccct cagctgatgg gcatcattcc ccggatcgct
     2221 cgagacatct tcaaccacat ctactccatg gatgagaacc ttgaattcca cattaaggta
     2281 tcttacttcg agatttacct ggataagatc cgtgaccttt tggatgtgac caagacgaac
     2341 ctgtccgtgc atgaggacaa aaaccgggtg ccgtttgtca agggttgtac cgaacgcttt
     2401 gtgtccagcc cagaggagat tctggatgtg atcgatgagg ggaagtccaa ccgtcacgta
     2461 gctgtcacca acatgaacga gcacagttct cggagccaca gcatcttcct catcaacatc
     2521 aagcaggaga acgtagagac cgagcagaag ctcagcggga agctgtacct cgtggatctg
     2581 gccggaagcg agaaggtcag caagacaggg gcagagggag ccgttctgga cgaggcaaag
     2641 aatatcaaca agtcgctgtc ggccctgggg aacgtgatct ctgcactggc agagggcacc
     2701 aaaagctacg tgccgtaccg cgacagcaaa atgacgagga ttctccagga ctctctggga
     2761 gggaactgca ggactaccat gttcatctgc tgctcgccgt ccagctacaa tgacgcagag
     2821 accaagtcca cgctcatgtt tggacagcgg gcgaagacca tcaagaacac tgcctcagtg
     2881 aatctggagc tgactgctga gcagtggaag aagaagtatg agaaggagaa ggagaagacc
     2941 aaggcccaga aggagacaat tgcgaagcta gaggctgagc ttagccggtg gcgcaatgga
     3001 gagaatgtgc ctgagactga gcgcctggct ggagaggact cagctctggg agctgagctc
     3061 tgcgaggaga cccctgtgaa tgacaactca tccattgtgg tacgcatcgc acctgaggaa
     3121 aggcagaaat atgaggaaga gatccgccgt ctctacaagc agcttgatga caaggatgat
     3181 gagatcaacc agcagagcca gctcattgag aagctgaagc agcagatgct ggaccaggaa
     3241 gagctgctcg tgtccactcg gggagacaac gagaaggtcc agcgggagct tagccacctg
     3301 cagtccgaga acgatgctgc gaaggacgag gtgaaggaag tgctgcaggc cctagaggag
     3361 ctggcggtca actacgacca gaagtcccag gaggtggagg agaagagcca gcagaaccag
     3421 ctgctggtgg acgagctgtc ccagaaagtg gccaccatgc tgtccctgga gtccgagcta
     3481 cagcggctcc aggaggtcag tggacaccag cgaaagcgga tcgctgaggt gctgaatggg
     3541 ctgatgaggg acctgagtga gttcagtgtc atcgtgggca acggcgagat taagctgccc
     3601 tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     3661 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3721 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3781 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3841 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     3901 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3961 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4021 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4081 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4141 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4201 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4261 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4321 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4381 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4441 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4501 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4561 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4621 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4681 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     4741 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4801 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4861 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4921 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4981 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgtgcgaagt
     5041 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5101 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5161 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5221 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     5281 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5341 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5401 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5461 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5521 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5581 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5641 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5701 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5761 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5821 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     5881 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5941 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6001 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6061 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6121 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6181 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6241 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6301 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6361 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6421 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6481 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6541 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6601 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6661 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6721 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6781 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6841 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6901 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6961 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7021 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7081 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7141 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7201 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7261 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7321 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7381 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7441 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7501 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7561 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7621 tgagaatagt gtatgcggc
//
