LOCUS       sequence-441237-        5830 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressed sfGFP fused human KIF1Bß(1-721) in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5830)
  TITLE     Characterizing Human KIF1Bß Motor Activity by Single-Molecule 
            Motility Assays and Caenorhabtidis elegans Genetics
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5830)
  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            "Unpublished"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: pSN667
FEATURES             Location/Qualifiers
     source          1..5830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      190..778
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     679..698
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  913..1136
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1203..1736)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1906..1911,1912..1928,1929..1934))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(2005..2038)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2115..2206
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2131..2148
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             join(4452..4454,4455..5165)
                     /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(5178..5201,5202..5237,5238..5261)
                     /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    5410..5544
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5447..5466)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5501..5520
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  5665..5830
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 accggttgac ttgggtcaac tgtcagacca agtttactca tatatacttt agattgattt
       61 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
      121 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
      181 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
      241 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      301 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
      361 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      421 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      481 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      541 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      601 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      661 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      721 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      781 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      841 ctttcctgcg ttatcccctg attgacttgg gtcgctcttc ctgtggatgc gcagatgccc
      901 tgcgtaagcg ggtgtgggcg gacaataaag tcttaaactg aacaaaatag atctaaacta
      961 tgacaataaa gtcttaaact agacagaata gttgtaaact gaaatcagtc cagttatgct
     1021 gtgaaaaagc atactggact tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc
     1081 aaattgcccg tcgtattaaa gaggggcgtg gccaagggca tgtaaagact atattcgcgg
     1141 cgttgtgaca atttaccgaa caactccgcg gccgggaagc cgatctcggc ttgaacgaat
     1201 tgttaggtgg cggtacttgg gtcgatatca aagtgcatca cttcttcccg tatgcccaac
     1261 tttgtataga gagccactgc gggatcgtca ccgtaatctg cttgcacgta gatcacataa
     1321 gcaccaagcg cgttggcctc atgcttgagg agattgatga gcgcggtggc aatgccctgc
     1381 ctccggtgct cgccggagac tgcgagatca tagatataga tctcactacg cggctgctca
     1441 aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca
     1501 agcgcgatga atgtcttact acggagcaag ttcccgaggt aatcggagtc cggctgatgt
     1561 tgggagtagg tggctacgtc tccgaactca cgaccgaaaa gatcaagagc agcccgcatg
     1621 gatttgactt ggtcagggcc gagcctacat gtgcgaatga tgcccatact tgagccacct
     1681 aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctcca taacatcaaa
     1741 catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga ctgtacaaaa
     1801 aaacagtcat aacaagccat gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc
     1861 aaggttctgg accagttgcg tgagcgcata cgctacttgc attacagttt acgaaccgaa
     1921 caggcttatg tcaactgggt tcgtgccttc atccgtttcc acggtgtgcg tcacccggca
     1981 accttgggca gcagcgaagt cgccataact tcgtatagca tacattatac gaagttatct
     2041 gtaactataa cggtcctaag gtagcgagtt taaacactag tatcgattcg cgacctactc
     2101 cggaatatta atagatcatg gagataatta aaatgataac catctcgcaa ataaataagt
     2161 attttactgt tttcgtaaca gttttgtaat aaaaaaacct ataaatattc cggattattc
     2221 ataccgtccc accatcgggc gcggatcccg gtccgaagcg cgcggaattc atgtccggcg
     2281 cttcagtgaa ggtggccgtg cgtgttcgtc ctttcaactc ccgtgaaacc tccaaagaat
     2341 ccaagtgcat catccagatg cagggcaact ccacctccat catcaaccct aagaacccca
     2401 aagaggctcc caagtccttc tccttcgact actcctactg gtcccacacc tcacctgagg
     2461 acccctgctt cgcttctcag aaccgtgtgt acaacgacat cggcaaagag atgctgctgc
     2521 acgctttcga gggttacaac gtgtgcatct tcgcttacgg ccagaccggt gctggaaagt
     2581 cctacaccat gatgggcaag caagaggaat cccaggctgg tatcatccct cagctgtgcg
     2641 aggaactgtt cgagaagatc aacgacaact gcaacgaaga gatgtcctac tccgtcgagg
     2701 tgtcctacat ggaaatctac tgcgagcgtg tgcgtgacct gctgaacccc aagaacaagg
     2761 gcaacctgcg tgtgcgcgaa caccctctgt tgggacctta cgtcgaggac ctgtccaagc
     2821 tggctgtgac ctcctacacc gatatcgctg acctgatgga cgctggcaac aaggctcgta
     2881 ccgtggctgc taccaacatg aacgagacat cctctcgttc ccacgctgtg ttcaccatcg
     2941 tgttcaccca gaagaagcac gacaacgaga ctaacctgtc caccgagaag gtgtccaaga
     3001 tctccctggt ggacctggct ggttctgagc gtgctgactc taccggtgct aagggcaccc
     3061 gtctgaaaga gggtgctaac atcaacaagt ccctgaccac tctgggcaaa gtgatctccg
     3121 ctctggctga ggtgtccaag aagaagaaaa agaccgactt catcccctac cgtgactccg
     3181 tgctgacctg gctcttgcgt gaaaacctcg gtggaaactc ccgtaccgct atggtggctg
     3241 ctctgtcccc tgctgacatc aactacgacg agactctgtc taccctgcgt tacgctgacc
     3301 gtgctaagca gatcaagtgc aacgctgtga tcaacgagga ccccaacgct aagctcgtgc
     3361 gcgagctgaa agaagaagtg acccgcctga aggacctgct gcgtgctcaa ggactgggcg
     3421 acatcatcga cacctctatg ggttccctga cctcctctcc atcttcatgc tccctgtcct
     3481 ctcaagtggg actgacctcc gtgacctcca tccaagagcg tatcatgtct acccctggtg
     3541 gcgaggaagc tatcgagcgc ctgaaagagt ccgagaagat tatcgccgag ctgaacgaaa
     3601 cctgggaaga gaagctgcgc aagaccgagg ctatccgtat ggaacgtgaa gctctgctgg
     3661 ctgagatggg tgtcgctatc cgtgaagatg gtggcaccct gggagtgttc tccccaaaga
     3721 aaacccctca cctggtcaac ctgaacgagg atcccctgat gtccgagtgc ctgctgtact
     3781 acatcaagga cggtatcacc cgtgtcggcc aggctgatgc tgaacgtcgt caagacatcg
     3841 tgctgtccgg tgctcatatc aaagaggaac actgcatctt ccgttccgag cgttccaact
     3901 ccggcgaagt gatcgtgacc ctggaacctt gcgagcgctc cgagacttac gtgaacggca
     3961 agcgtgtgtc ccagcctgtg caactgagat ccggcaaccg tatcatcatg ggcaagaacc
     4021 acgtgttccg tttcaaccat cctgagcagg ctcgtgctga gcgtgaaaag accccttccg
     4081 ctgagactcc ttccgagcct gtggactgga ctttcgctca gcgtgaactg ctggaaaagc
     4141 agggtatcga catgaagcaa gagatggaaa agcgcctgca agaaatggaa attctgtaca
     4201 agaaagagaa agaggaagcc gacctgctgt tggagcagca gcgtctggac tacgagtcta
     4261 agctgcaggc tctgcagaag caggtcgaga ctcgttccct ggctgctgag actaccgagg
     4321 aagaagagga agaggaagaa gtcccctgga cacagcacga gttcgagctg gctcagtggg
     4381 ctttccgcaa gtggaagtcc caccagttca cttccctgcg cgacctgttg tgggggcggc
     4441 cgctttcgaa tatgagtaaa ggtgaggaac tctttacagg agttgtaccc atactcgttg
     4501 agttggacgg tgatgtaaat ggtcataagt tttcggtcag gggtgagggc gagggtgacg
     4561 caactaacgg taagctgact ttgaagttta tctgtaccac gggaaaactt cccgtgccgt
     4621 ggccaacttt ggtaacaaca cttacatacg gtgtgcaatg ctttagtcgc tacccggatc
     4681 acatgaaaag gcacgatttt ttcaaatctg ccatgcctga gggttatgtg caagagagaa
     4741 ctatttcgtt caaagacgac ggaacgtaca agacacgcgc cgaagtgaaa tttgagggtg
     4801 atacccttgt caatcgtata gaacttaaag gaattgactt caaggaagac ggcaacatct
     4861 tgggacacaa actggaatat aattttaact cgcacaacgt gtatatcaca gcagacaaac
     4921 agaaaaacgg aatcaaagca aacttcaaaa tacgccacaa cgttgaagat ggaagcgtcc
     4981 agctggctga tcattaccag caaaacaccc ccattggtga tggaccggta ttgttgcccg
     5041 ataatcatta tctctcaacg cagtctgtac tcagtaagga tcctaacgaa aagagggatc
     5101 acatggttct tctcgagttc gtaaccgcag ctggtatcac gcacggcatg gacgaactct
     5161 acaaaggcag tggcagttgg tcgcatcctc aattcgagaa gggcggcggc agtggcggcg
     5221 gcagtggcgg cagtgcctgg agtcatccac aatttgagaa gtaaagcctg cagtctcgac
     5281 aagcttgtcg agaagtacta gaggatcata atcagccata ccacatttgt agaggtttta
     5341 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     5401 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5461 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5521 aatgtatctt atcatgtctg gatctgatca ctgcttgagc ctagaagatc cggctgctaa
     5581 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tatcataacc
     5641 cctagggtat acccatctaa ttggaaccag ataagtgaaa tctagttcca aactattttg
     5701 tcatttttaa ttttcgtatt agcttacgac gctacaccca gttcccatct attttgtcac
     5761 tcttccctaa ataatcctta aaaactccat ttccacccct cccagttccc aactattttg
     5821 tccgcccaca
//
