LOCUS       sequence-435685-        9001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of CLIPf-KIF5A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9001)
  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 9001)
  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
FEATURES             Location/Qualifiers
     source          1..9001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     474..493
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    778..899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(815..834)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     869..888
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(948..964)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(948..964)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(961..983)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    972..988
                     /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(996..1002,1003..1020,1021..1026))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1041..1062
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1179..1196)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1350..1938)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1430..1449)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2109..2900,2901..2969))
                     /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     2732..2751
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2970..3074)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3149..3168)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3340..3719
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3720..3923
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3874..3894
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3968..3987
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3968..3986
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7697..7714)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7703..7927
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7973..8401
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8060..8079)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8270..8291
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8410..8430)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8415..8744
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8595..8730
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8657..8676
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             join(8811..9001,1..604)
                     /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(8865..8884)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
       61 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
      121 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
      181 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
      241 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
      301 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
      361 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
      421 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
      481 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
      541 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
      601 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
      661 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
      721 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
      781 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
      841 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
      901 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
      961 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     1021 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     1081 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     1141 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     1201 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1261 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1321 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1381 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1441 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1501 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1561 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1621 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1681 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1741 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     1801 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1861 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1921 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1981 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2041 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2101 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2161 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     2221 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     2281 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     2341 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     2401 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     2461 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     2521 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     2581 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     2641 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     2701 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     2761 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     2821 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     2881 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     2941 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3001 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3061 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     3121 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3181 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     3241 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     3301 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
     3361 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     3421 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     3481 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     3541 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     3601 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     3661 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     3721 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     3781 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     3841 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     3901 gggaggtcta tataagcaga gctctctggc taactagaga acccactgct tactggctta
     3961 tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta aacttaagct
     4021 tggtaccatg gacaaggact gcgagatgaa gcggaccaca ctggatagcc ctctgggcaa
     4081 gctggaactg agcggctgtg aacagggcct gcaccggatc atctttctcg gcaagggaac
     4141 ctctgccgcc gatgctgttg aagtgcctgc tcctgctgct gtgctcggag gacctgaacc
     4201 tctgattcag gccaccgcct ggctgaacgc ctactttcat cagcccgagg ccatcgagga
     4261 gttccccgtt cctgctctgc atcaccccgt gttccagcaa gaaagcttca ccagacaggt
     4321 gctgtggaag ctgctgaagg tggtcaagtt cggcgaagtg atcagcgagt ctcacctggc
     4381 cgctctcgtg ggaaatccag ctgctacagc cgccgtgaac acagccctgg atggaaaccc
     4441 tgtgcctatt ctgatcccct gccacagagt ggtgcagggc gattctgatg tgggccctta
     4501 tcttggaggc ctggccgtga aagaatggct gctggcccac gaaggacaca gactgggaaa
     4561 acctggcctc ggaggatcca tggcggagac taacaacgaa tgcagcatca aggtgctttg
     4621 ccgatttcgg cccctgaacc aggccgagat tctgcggggg gacaagttca tccccatttt
     4681 ccaaggggac gacagcgtca ttattggggg aaagccatat gtctttgacc gcgtcttccc
     4741 cccaaacacc actcaggagc aggtttacca cgcctgtgcc atgcagatcg tcaaagacgt
     4801 ccttgctggt tacaatggca caatcttcgc ttatggacag acatcctcag ggaaaacgca
     4861 taccatggag gggaagctgc acgaccctca gctgatgggc atcattcccc ggatcgctcg
     4921 agacatcttc aaccacatct actccatgga tgagaacctt gaattccaca ttaaggtatc
     4981 ttacttcgag atttacctgg ataagatccg tgaccttttg gatgtgacca agacgaacct
     5041 gtccgtgcat gaggacaaaa accgggtgcc gtttgtcaag ggttgtaccg aacgctttgt
     5101 gtccagccca gaggagattc tggatgtgat cgatgagggg aagtccaacc gtcacgtagc
     5161 tgtcaccaac atgaacgagc acagttctcg gagccacagc atcttcctca tcaacatcaa
     5221 gcaggagaac gtagagaccg agcagaagct cagcgggaag ctgtacctcg tggatctggc
     5281 cggaagcgag aaggtcagca agacaggggc agagggagcc gttctggacg aggcaaagaa
     5341 tatcaacaag tcgctgtcgg ccctggggaa cgtgatctct gcactggcag agggcaccaa
     5401 aagctacgtg ccgtaccgcg acagcaaaat gacgaggatt ctccaggact ctctgggagg
     5461 gaactgcagg actaccatgt tcatctgctg ctcgccgtcc agctacaatg acgcagagac
     5521 caagtccacg ctcatgtttg gacagcgggc gaagaccatc aagaacactg cctcagtgaa
     5581 tctggagctg actgctgagc agtggaagaa gaagtatgag aaggagaagg agaagaccaa
     5641 ggcccagaag gagacaattg cgaagctaga ggctgagctt agccggtggc gcaatggaga
     5701 gaatgtgcct gagactgagc gcctggctgg agaggactca gctctgggag ctgagctctg
     5761 cgaggagacc cctgtgaatg acaactcatc cattgtggta cgcatcgcac ctgaggaaag
     5821 gcagaaatat gaggaagaga tccgccgtct ctacaagcag cttgatgaca aggatgatga
     5881 gatcaaccag cagagccagc tcattgagaa gctgaagcag cagatgctgg accaggaaga
     5941 gctgctcgtg tccactcggg gagacaacga gaaggtccag cgggagctta gccacctgca
     6001 gtccgagaac gatgctgcga aggacgaggt gaaggaagtg ctgcaggccc tagaggagct
     6061 ggcggtcaac tacgaccaga agtcccagga ggtggaggag aagagccagc agaaccagct
     6121 gctggtggac gagctgtccc agaaagtggc caccatgctg tccctggagt ccgagctaca
     6181 gcggctccag gaggtcagtg gacaccagcg aaagcggatc gctgaggtgc tgaatgggct
     6241 gatgagggac ctgagtgagt tcagtgtcat cgtgggcaac ggcgagatta agctgcccgt
     6301 ggagatcagt ggggccatcg aggaggagtt cacggtggcc cggctctaca tcagcaagat
     6361 caagtcggag gtgaagtccg tggttaagcg atgtcggcag ctggagaacc tccaggtgga
     6421 gtgtcatcgc aagatggagg tgaccggtag ggagctgtca tcttgccaac tgctcatctc
     6481 acagcatgag gccaagatcc gttcactcac ggagtacatg cagactgtgg agttgaagaa
     6541 acggcacctg gaagagtcct acgactccct gagcgatgag cttgccaggc tccaggcgca
     6601 cgaaactgta cacgaggtag ctctgaaaga caaggagcca gacacacagg acgcggagga
     6661 ggtgaagaag gccctggaac tacagatgga gaatcaccgt gaggcccatc accggcagct
     6721 ggcccgcctc cgggatgaga ttaatgagaa acagaaaacc attgatgagc tgaaagacct
     6781 gaaccagaag ctccagttag agctggagaa gcttcaggcc gactatgaga ggctgaagaa
     6841 tgaagagaac gagaagagcg ccaagctcca ggagctgaca tttctgtatg agcgacatga
     6901 gcagtccaag caggacctca aggggctgga ggagacagtt gcccgtgaac tccagaccct
     6961 ccacaacctt cgcaagctgt tcgttcaaga cgtcacgact cgagtcaaga aaagtgcaga
     7021 aatggagccc gaggacagtg gggggattca ttcccaaaag cagaagatct cctttcttga
     7081 gaacaacctg gaacagctta caaaggttca caaacagctg gtacgtgaca atgcagatct
     7141 gcgttgtgag cttcctaaat tggaaaaacg acttcgggct acggctgaga gagttaaggc
     7201 cctggagggt gcactgaagg aggccaagga gggcgctatg aaggacaagc gtagatacca
     7261 gcaggaggtg gaccgcatca aagaagccgt gcggtacaag agctccggca agcggggcca
     7321 ttctgcccag atcgctaagc ctgtgaggcc tggccactat cctgcctcct cacccaccaa
     7381 cccctacggc acccggagcc ccgagtgtat cagctacacc aacaacctct tccagaacta
     7441 ccagaacctg cacctgcagg ctgcgcctag ctccacttca gatatgtact ttgccagcag
     7501 cggagccaca tctgttgccc ccttggcttc ctaccagaag gccaacatgg acaatggaaa
     7561 tgccacagat atcaacgaca acaggagtga cctgccgtgt ggctatgagg ctgaggacca
     7621 ggccaagctt ttccctctcc accaagagac agcagccagc tcgagtctag agggcccgtt
     7681 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     7741 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     7801 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     7861 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     7921 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     7981 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     8041 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     8101 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     8161 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     8221 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     8281 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     8341 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     8401 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     8461 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     8521 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     8581 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     8641 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     8701 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     8761 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     8821 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     8881 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     8941 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     9001 c
//
