LOCUS       sequence-440846-        8996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressed sfGFP fused human KIF1Bß (1770 a.a. isoform) in insect 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8996)
  TITLE     Characterizing Human KIF1Bß Motor Activity by Single-Molecule 
            Motility Assays and Caenorhabtidis elegans Genetics
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8996)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pSN668
FEATURES             Location/Qualifiers
     source          1..8996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      73..661
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     562..581
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  796..1019
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1086..1619)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1808..1813,1814..1830,1831..1836))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(1907..1940)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2017..2108
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2033..2050
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             join(7501..7503,7504..8214)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             join(8227..8250,8251..8286,8287..8310)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="WSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     polyA_signal    8459..8593
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8496..8515)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8550..8569
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  8714..8879
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
       61 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
      121 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
      181 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
      241 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
      301 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
      361 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
      421 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
      481 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
      541 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
      601 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
      661 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
      721 gttctttcct gcgttatccc ctgattgact tgggtcgctc ttcctgtgga tgcgcagatg
      781 ccctgcgtaa gcgggtgtgg gcggacaata aagtcttaaa ctgaacaaaa tagatctaaa
      841 ctatgacaat aaagtcttaa actagacaga atagttgtaa actgaaatca gtccagttat
      901 gctgtgaaaa agcatactgg acttttgtta tggctaaagc aaactcttca ttttctgaag
      961 tgcaaattgc ccgtcgtatt aaagaggggc gtggccaagg gcatgtaaag actatattcg
     1021 cggcgttgtg acaatttacc gaacaactcc gcggccggga agccgatctc ggcttgaacg
     1081 aattgttagg tggcggtact tgggtcgata tcaaagtgca tcacttcttc ccgtatgccc
     1141 aactttgtat agagagccac tgcgggatcg tcaccgtaat ctgcttgcac gtagatcaca
     1201 taagcaccaa gcgcgttggc ctcatgcttg aggagattga tgagcgcggt ggcaatgccc
     1261 tgcctccggt gctcgccgga gactgcgaga tcatagatat agatctcact acgcggctgc
     1321 tcaaacttgg gcagaacgta agccgcgaga gcgccaacaa ccgcttcttg gtcgaaggca
     1381 gcaagcgcga tgaatgtctt actacggagc aagttcccga ggtaatcgga gtccggctga
     1441 tgttgggagt aggtggctac gtctccgaac tcacgaccga aaagatcaag agcagcccgc
     1501 atggatttga cttggtcagg gccgagccta catgtgcgaa tgatgcccat acttgagcca
     1561 cctaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct gcgtaacatc
     1621 gttgctgctc cataacatca aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc
     1681 ccgaggcata gactgtacaa aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc
     1741 gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca tacgctactt
     1801 gcattacagt ttacgaaccg aacaggctta tgtcaactgg gttcgtgcct tcatccgttt
     1861 ccacggtgtg cgtcacccgg caaccttggg cagcagcgaa gtcgccataa cttcgtatag
     1921 catacattat acgaagttat ctgtaactat aacggtccta aggtagcgag tttaaacact
     1981 agtatcgatt cgcgacctac tccggaatat taatagatca tggagataat taaaatgata
     2041 accatctcgc aaataaataa gtattttact gttttcgtaa cagttttgta ataaaaaaac
     2101 ctataaatat tccggattat tcataccgtc ccaccatcgg gcgcggatcc cggtccgaag
     2161 cgcgcggaat tcatgtccgg cgcttcagtg aaggtggccg tgcgtgttcg tcctttcaac
     2221 tcccgtgaaa cctccaaaga atccaagtgc atcatccaga tgcagggcaa ctccacctcc
     2281 atcatcaacc ctaagaaccc caaagaggct cccaagtcct tctccttcga ctactcctac
     2341 tggtcccaca cctcacctga ggacccctgc ttcgcttctc agaaccgtgt gtacaacgac
     2401 atcggcaaag agatgctgct gcacgctttc gagggttaca acgtgtgcat cttcgcttac
     2461 ggccagaccg gtgctggaaa gtcctacacc atgatgggca agcaagagga atcccaggct
     2521 ggtatcatcc ctcagctgtg cgaggaactg ttcgagaaga tcaacgacaa ctgcaacgaa
     2581 gagatgtcct actccgtcga ggtgtcctac atggaaatct actgcgagcg tgtgcgtgac
     2641 ctgctgaacc ccaagaacaa gggcaacctg cgtgtgcgcg aacaccctct gttgggacct
     2701 tacgtcgagg acctgtccaa gctggctgtg acctcctaca ccgatatcgc tgacctgatg
     2761 gacgctggca acaaggctcg taccgtggct gctaccaaca tgaacgagac atcctctcgt
     2821 tcccacgctg tgttcaccat cgtgttcacc cagaagaagc acgacaacga gactaacctg
     2881 tccaccgaga aggtgtccaa gatctccctg gtggacctgg ctggttctga gcgtgctgac
     2941 tctaccggtg ctaagggcac ccgtctgaaa gagggtgcta acatcaacaa gtccctgacc
     3001 actctgggca aagtgatctc cgctctggct gaggtgtcca agaagaagaa aaagaccgac
     3061 ttcatcccct accgtgactc cgtgctgacc tggctcttgc gtgaaaacct cggtggaaac
     3121 tcccgtaccg ctatggtggc tgctctgtcc cctgctgaca tcaactacga cgagactctg
     3181 tctaccctgc gttacgctga ccgtgctaag cagatcaagt gcaacgctgt gatcaacgag
     3241 gaccccaacg ctaagctcgt gcgcgagctg aaagaagaag tgacccgcct gaaggacctg
     3301 ctgcgtgctc aaggactggg cgacatcatc gacacctcta tgggttccct gacctcctct
     3361 ccatcttcat gctccctgtc ctctcaagtg ggactgacct ccgtgacctc catccaagag
     3421 cgtatcatgt ctacccctgg tggcgaggaa gctatcgagc gcctgaaaga gtccgagaag
     3481 attatcgccg agctgaacga aacctgggaa gagaagctgc gcaagaccga ggctatccgt
     3541 atggaacgtg aagctctgct ggctgagatg ggtgtcgcta tccgtgaaga tggtggcacc
     3601 ctgggagtgt tctccccaaa gaaaacccct cacctggtca acctgaacga ggatcccctg
     3661 atgtccgagt gcctgctgta ctacatcaag gacggtatca cccgtgtcgg ccaggctgat
     3721 gctgaacgtc gtcaagacat cgtgctgtcc ggtgctcata tcaaagagga acactgcatc
     3781 ttccgttccg agcgttccaa ctccggcgaa gtgatcgtga ccctggaacc ttgcgagcgc
     3841 tccgagactt acgtgaacgg caagcgtgtg tcccagcctg tgcaactgag atccggcaac
     3901 cgtatcatca tgggcaagaa ccacgtgttc cgtttcaacc atcctgagca ggctcgtgct
     3961 gagcgtgaaa agaccccttc cgctgagact ccttccgagc ctgtggactg gactttcgct
     4021 cagcgtgaac tgctggaaaa gcagggtatc gacatgaagc aagagatgga aaagcgcctg
     4081 caagaaatgg aaattctgta caagaaagag aaagaggaag ccgacctgct gttggagcag
     4141 cagcgtctgg actacgagtc taagctgcag gctctgcaga agcaggtcga gactcgttcc
     4201 ctggctgctg agactaccga ggaagaagag gaagaggaag aagtcccctg gacacagcac
     4261 gagttcgagc tggctcagtg ggctttccgc aagtggaagt cccaccagtt cacttccctg
     4321 cgcgacctgt tgtggggaaa cgctgtgtac ttgaaagagg ctaacgctat ctccgtggaa
     4381 ctgaagaaga aggtccagtt ccagttcgtg ctgctgaccg atactctgta cagccctctg
     4441 cctcctgagc tgctgcctac cgaaatggaa aagacccacg aggaccgtcc ttttccacgc
     4501 actgtggtgg ctgtggaagt gcaggacctg aagaacggtg ctacccacta ctggtctttg
     4561 gagaagctca agcagaggct ggacctgatg cgcgagatgt acgatcgtgc tggcgaaatg
     4621 gcttccagcg ctcaggacga gtctgaaacc accgtgaccg gttctgaccc cttctacgac
     4681 cgtttccact ggttcaagct cgtgggcagc tcccctatct tccacggttg cgtgaacgag
     4741 cgtctggctg acagaacccc ttctcctacc ttctcaaccg ctgactccga catcactgag
     4801 ctggctgacg agcagcagga cgagatggaa gatttcgacg acgaggcttt cgtggacgac
     4861 gctggatctg acgctggtac tgaggaaggt tccgacctgt tctccgacgg tcacgaccca
     4921 ttctatgaca ggtccccatg gttcatcctc gtcggtcgtg ctttcgtgta cctgtctaac
     4981 ctgctctacc ccgtgcctct gatccaccgt gtggctatcg tgtctgagaa gggcgaagtg
     5041 cgcggtttct tgcgtgtggc tgtgcaggct atcgctgctg acgaagaagc tcccgactac
     5101 ggttccggta tccgtcagtc cggaaccgct aagatctctt tcgacaacga atacttcaac
     5161 cagtccgact tctcctccgt ggctatgacc cgttccggcc tgtctctgga agaactgcgt
     5221 atcgtggaag gccagggcca gtcctccgaa gttatcaccc ctcctgagga aatctcccgc
     5281 atcaacgacc tggacctcaa gtcctccact ctgctggacg gcaagatggt catggaaggc
     5341 ttctccgaag agatcggaaa ccacctgaag ctgggttccg ctttcacctt ccgtgtgacc
     5401 gtcctgcagg cctctggtat cctgcctgag tacgctgaca tcttctgcca attcaacttc
     5461 ctgcaccgtc acgacgaagc cttctctacc gagcctctga agaacaacgg tcgtggttcc
     5521 cctctggctt tctaccacgt gcagaacatt gctgtcgaga tcaccgagtc cttcgtggat
     5581 tacatcaaga ccaagcctat cgtgttcgag gtgttcggtc actaccagca gcacccactg
     5641 cacttgcaag gccaagagct gaacagccct cctcagcctt gccgtcgttt cttccctcct
     5701 ccaatgcctc tgtctaagcc tgtgcctgcc accaagctga acaccatgtc caagacttct
     5761 ctgggccaga gcatgtctaa gtacgacctg ctcgtttggt tcgagatctc tgagctggaa
     5821 cccaccggcg agtacatccc tgctgtggtg gatcacactg ctggcctgcc ttgtcagggc
     5881 actttcctgt tgcaccaggg tatccagcgc cgtatcaccg tcaccatcat ccacgagaag
     5941 ggttccgagc tgcactggaa ggacgtgcgt gaactcgtcg tgggtcgtat ccgtaacaag
     6001 cccgaggtgg acgaagctgc tgtggacgct atcctgagcc tgaacatcat ctccgctaag
     6061 tacttgaagt cgtcccacaa ctcctccagg accttctacc gtttcgaggc tgtgtgggac
     6121 tcctctctgc acaactctct gctgctcaac cgtgtcaccc cttacggcga aaagatctac
     6181 atgaccctgt ccgcttacct cgagctggac cactgcatcc agcctgctgt catcaccaag
     6241 gacgtgtgta tggtgttcta ctcccgcgac gccaagatca gccctccacg ttctctccgt
     6301 tctctgttcg gttccggcta ctccaagtct cccgactcca accgtgtgac tggtatctac
     6361 gagctgtccc tgtgcaagat gtccgacacc ggttctcccg gaatgcagcg taggcgtagg
     6421 aagatcctgg atacctccgt cgcttacgtg cgtggcgaag agaacttggc tggatggcgt
     6481 cctaggggcg actccttgat cctggaacac cagtgggaac tcgagaagct ggaactgctc
     6541 cacgaggtcg aaaagaccag gcacttcttg ctgctgcgcg agaggctggg agactctatc
     6601 cccaagtctc tgtccgactc actgtcccct tctctgtcct ccggtactct ctctacctcc
     6661 acctctatct ccagccagat ctccaccacc accttcgagt ccgctatcac cccatccgag
     6721 tcctctggtt acgactctgg cgacatcgag tcactggtcg accgcgagaa agaactggct
     6781 accaagtgcc tgcaactgct gacccacacc ttcaaccgcg agttctctca ggtgcacggt
     6841 tccgtgtctg actgcaagct gtctgacatc tctcccatcg gacgtgaccc ctctgagtcc
     6901 tccttcagct ccgctactct gactccctcc tcaacttgcc cctctctggt ggactcccgt
     6961 tctaactccc tggaccaaaa gactcccgag gctaactccc gcgctagctc tccttgtcct
     7021 gagttcgaac aattccagat cgtgcccgct gtggaaaccc catacttggc tcgcgctggc
     7081 aagaacgagt tcctgaacct ggtgcctgac atcgaggaaa tccgtcctag ctccgtggtt
     7141 tccaagaagg gttacctcca cttcaaagag cccctgtact ccaactgggc taagcacttc
     7201 gtggtcgtgc gtcgtcccta cgtgttcatc tacaactccg acaaggaccc cgtcgagcgc
     7261 ggtatcatca acttgtctac cgctcaggtg gaatactccg aggaccagca ggctatggtc
     7321 aagaccccta acaccttcgc tgtctgcacc aagcacaggg gcgtgctgtt gcaagctctg
     7381 aacgacaagg atatgaacga ctggctgtac gctttcaacc ctctgctcgc tggcaccatc
     7441 cgttccaagt tgtctcgtag gtgccccagc caatctaagt acgggcggcc gctttcgaat
     7501 atgagtaaag gtgaggaact ctttacagga gttgtaccca tactcgttga gttggacggt
     7561 gatgtaaatg gtcataagtt ttcggtcagg ggtgagggcg agggtgacgc aactaacggt
     7621 aagctgactt tgaagtttat ctgtaccacg ggaaaacttc ccgtgccgtg gccaactttg
     7681 gtaacaacac ttacatacgg tgtgcaatgc tttagtcgct acccggatca catgaaaagg
     7741 cacgattttt tcaaatctgc catgcctgag ggttatgtgc aagagagaac tatttcgttc
     7801 aaagacgacg gaacgtacaa gacacgcgcc gaagtgaaat ttgagggtga tacccttgtc
     7861 aatcgtatag aacttaaagg aattgacttc aaggaagacg gcaacatctt gggacacaaa
     7921 ctggaatata attttaactc gcacaacgtg tatatcacag cagacaaaca gaaaaacgga
     7981 atcaaagcaa acttcaaaat acgccacaac gttgaagatg gaagcgtcca gctggctgat
     8041 cattaccagc aaaacacccc cattggtgat ggaccggtat tgttgcccga taatcattat
     8101 ctctcaacgc agtctgtact cagtaaggat cctaacgaaa agagggatca catggttctt
     8161 ctcgagttcg taaccgcagc tggtatcacg cacggcatgg acgaactcta caaaggcagt
     8221 ggcagttggt cgcatcctca attcgagaag ggcggcggca gtggcggcgg cagtggcggc
     8281 agtgcctgga gtcatccaca atttgagaag taaagcctgc agtctcgaca agcttgtcga
     8341 gaagtactag aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     8401 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     8461 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     8521 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     8581 tcatgtctgg atctgatcac tgcttgagcc tagaagatcc ggctgctaac aaagcccgaa
     8641 aggaagctga gttggctgct gccaccgctg agcaataact atcataaccc ctagggtata
     8701 cccatctaat tggaaccaga taagtgaaat ctagttccaa actattttgt catttttaat
     8761 tttcgtatta gcttacgacg ctacacccag ttcccatcta ttttgtcact cttccctaaa
     8821 taatccttaa aaactccatt tccacccctc ccagttccca actattttgt ccgcccacaa
     8881 ccggttgact tgggtcaact gtcagaccaa gtttactcat atatacttta gattgattta
     8941 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcat
//
