LOCUS       sequence-263568-        6128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses worm Kinesin-3 Unc104 head and neck linker, dimerized 
            through kinesin-1 neck-coil and coil-1 to 560 and GFP tagged.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6128)
  AUTHORS   Hancock
  TITLE     Hancock Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6128)
  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            "Unpublished"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: Unc104_K560GFP
FEATURES             Location/Qualifiers
     source          1..6128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1576..1584
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     regulatory      1784..1793
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1790..1792,1793..1795,1796..2506)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1790..1792,1793..1795,1796..2506)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1835..1856)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2096..2115)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2443..2464
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2516..2533
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2556..2573
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2626..2644)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2640..2687
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2724..3179
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2811..2830)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3021..3042
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3205..3309
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3310..3378,3379..4170)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3528..3547)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4341..4929
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4830..4849
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5083..5100
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5115..5254
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5180..5199)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5340..5362
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5356..5547)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6056..6075
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6056..6074
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             6106..6128
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 tatacatata catatgtcat cggttaaagt agctgtacgt gttcgcccat tcaaccaacg
       61 ggaaatctcg aacacttcaa aatgtgtcct tcaagtaaat ggaaatacga cgacaataaa
      121 tggtcattca attaacaagg agaacttcag tttcaatttt gatcattcat attggtcgtt
      181 tgcgaggaat gacccacatt ttatcaccca aaaacaagta tatgaagagc tcggagttga
      241 aatgttggaa cacgcatttg aagggtataa tgtctgcatt tttgcatacg gtcaaacagg
      301 atcaggaaaa tcatatacaa tgatgggaaa agccaatgat ccagatgaaa tgggtataat
      361 tccacgtttg tgcaatgatt tatttgcacg aattgataat aacaatgata aagatgttca
      421 atattctgta gaggtatcgt atatggaaat ttattgtgaa cgagtaaaag atcttctgaa
      481 tcctaactct ggaggtaacc taagggttcg tgaacatcct ttacttggac cttacgtcga
      541 tgaccttacc aaaatggcag tttgttctta ccacgacatt tgcaatctga tggacgaagg
      601 aaataaagcg agaactgttg ctgcaacaaa tatgaattcg acatcatcaa gatcccatgc
      661 ggtattcaca atagtactca ctcaaaaaag acattgtgct gattctaatt tggatactga
      721 gaagcattca aaaatttctt tggttgattt ggcaggatcc gaaagagcca attctacagg
      781 agcggaaggt caacgactaa aagaaggagc aaatatcaac aagagcttga caacgttggg
      841 tcttgtaatc agtaaacttg cagaagaatc aacaaagaag aaaaagtcca acaaaggtgt
      901 gattccttat cgtgattctg tgctgacgtg gcttctgaga gaaaatctgg gaggaaattc
      961 gaaaactgcg atgctcgcgg cattatctcc agctgatatt aactttgatg aaacactgag
     1021 tactttgaga tatgccgata gagcgaaaca aattgtttgt caagcagttg ttaatgagga
     1081 tccaaatgca aagctagccg aggaatggaa gcgacgctat gaaaaggaga aggaaaagaa
     1141 cgcccgacta aagggtaagg tggagaagct tgagatcgag cttgcgcgct ggagagcggg
     1201 tgaaactgtt aaggcggagg agcaaatcaa catggaggat ctcatggagg caagcacgcc
     1261 caacctggaa gtggaggcag cacagacggc ggcggccgag gccgctttgg ccgcccagcg
     1321 aacggctctc gccaatatgt ccgcatcggt tgccgtgaac gagcaggcca ggctggctac
     1381 agagtgcgag cgtctctacc agcagctgga cgacaaggat gaggagatca atcagcagag
     1441 ccagtacgcc gagcagctca aggagcaggt gatggagcag gaggaactca tcgctaacgc
     1501 tcggcgtgag tatgagactt tgcagtcgga gatggcgcga attcaacagg agaacgagtc
     1561 cgccaaggaa gaggttaagg aggtgctcca agctctcgaa gagctgactg taaactacga
     1621 ccagaaatcc caggagatcg ataacaagaa caaggatatc gatgccctca acgaggagct
     1681 gcagcagaag cagtctgtgt tcaacgccgc ctccacagag ctacagcagc tcaagatgct
     1741 tgttccgcgt gggcccgtgc accgcctagg ggatccaccg gtcgccacca tggtgagcaa
     1801 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1861 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1921 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1981 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     2041 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     2101 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2161 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     2221 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     2281 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     2341 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     2401 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     2461 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagctcg agagccatca
     2521 tcatcatcat cactaagcgg ccgcactcga tcgagcacca ccaccaccac cactgagatc
     2581 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
     2641 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
     2701 ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2761 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2821 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2881 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     2941 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3001 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3061 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3121 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     3181 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3241 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3301 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3361 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3421 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3481 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3541 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3601 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3661 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3721 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3781 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3841 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3901 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3961 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4021 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4081 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4141 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4201 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4261 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4321 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4381 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4441 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     4501 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4561 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4621 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4681 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4741 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4801 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4861 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4921 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4981 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     5041 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     5101 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     5161 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     5221 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     5281 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5341 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     5401 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     5461 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     5521 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     5581 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     5641 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     5701 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     5761 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     5821 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     5881 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     5941 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     6001 catgcgcacc cgtggccagg acccaacgct gcccgagatc tcgatcccgc gaaattaata
     6061 cgactcacta tagggagacc acaacggttt ccctctagaa ataattttgt ttaactttaa
     6121 gaaggaga
//
