LOCUS       sequence-140258-        5601 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5601)
  AUTHORS   Woehlke G, Ruby AK, Hart CL, Ly B, Hom-Booher N, Vale RD
  TITLE     Microtubule interaction site of the kinesin motor.
  JOURNAL   Cell. 1997 Jul 25. 90(2):207-16.
  PUBMED    9244295
REFERENCE   2  (bases 1 to 5601)
  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: "Cell. 1997 
            Jul 25. 90(2):207-16."
COMMENT     Sequence Label: pET17_K560_GFP_His
FEATURES             Location/Qualifiers
     source          1..5601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(28..75)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     71..89
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(150..167)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(168..881)
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             complement(168..881)
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(208..228)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     823..851
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             2575..2597
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2628..2647)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2629..2647)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3156..3347
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3341..3363)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3449..3588
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3504..3523
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3603..3620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3774..4362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3854..3873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4533..5324,5325..5393))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5156..5175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5394..5498)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5566..5584
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttatccggat atagttcctc ctttcagcaa aaaacccctc aagacccgtt tagaggcccc
       61 aaggggttat gctagttatt gctcagcggt ggcagcagcc aactcagctt cctttcgggc
      121 tttgttagca gccggatctg ctcgagttaa tggtgatgat ggtgatgttt gtatagttca
      181 tccatgccat gtgtaatccc agcagctgtt acaaactcaa gaaggaccat gtggtctctc
      241 ttttcgttgg gatctttcga aagggcagat tgtgtggaca ggtaatggtt gtctggtaaa
      301 aggacagggc catcgccaat tggagtattt tgttgataat ggtctgctag ttgaacgctt
      361 ccatcttcaa tgttgtgtct aattttgaag ttaactttga ttccattctt ttgtttgtct
      421 gccatgatgt atacattgtg tgagttatag ttgtattcca atttgtgtcc aagaatgttt
      481 ccatcttctt taaaatcaat accttttaac tcgattctat taacaagggt atcaccttca
      541 aacttgactt cagcacgtgt cttgtagttc ccgtcatctt tgaaaaatat agttctttcc
      601 tgtacataac cttcgggcat ggcactcttg aaaaagtcat gctgtttcat atgatctggg
      661 tatcttgaaa agcattgaac accataagtc agagtagtga caagtgttgg ccatggaaca
      721 ggtagttttc cagtagtgca aataaattta agggtaagtt ttccgtatgt tgcatcacct
      781 tcaccctctc cactgacaga aaatttgtgc ccattaacat caccatctaa ttcaacaaga
      841 attgggacaa ctccagtgaa aagttcttct cctttactca tggtaccttt tagtaaagat
      901 gccatcatct cagctgctcg ttttttctgg tggttggtca tttccttaag tttctgaagc
      961 tcagcatcta tactcgctaa agttgccgat ttctgattca attcatcact aagcaattca
     1021 tattccttag ttttgtcttc aacttcctga gacttctgat cataattgac agcaagttct
     1081 tctagggcct gtaaaacttc tttcacttct tctttagagg catcattttc tgcttgaagg
     1141 cgattcagct cagcttgcat attgtcttga tcccttctgg tagatgccaa aagctcctcc
     1201 tgatccaaca tttgcgtctt cagcttctct accagttgac tttgctggtt aatttcttca
     1261 tccttgtcat caagctgttt gtataattta gcaatttctt cttcacactt tcttctttca
     1321 gcatcagtaa aatttcctat aactccaatt gcggttgctg gtttatcatt ggtaagagta
     1381 atatctttat ccactgtgaa agcttccaag ttggctttct ctttgtcaaa ctgttcatca
     1441 ataggcaccg tctccccatt acgccatctg ttgagctcat tttcaagcca ctgaatagtg
     1501 ttccgcagga tcttattttt ttctttttct ttttcatact tctttttcca ctgttctgca
     1561 gttaactcca cattgacaca aactgtgttc ttaattgttt tggccctttg gccaaataag
     1621 agtgtagatt ttgtttcaga ctcattgtat gatgatggag agcagcaaat tacaatagtg
     1681 gttctacagt tgccacctaa tgaatcttga aggattcttg tcattttact atctcgatat
     1741 ggaacataag tactaccctc agccaaagca gaaataacat ttccaagagc agaaagtgac
     1801 ttgttgatgt ttttagcttc atccagcaca gcaccttcag ctccagtttt actaaccttt
     1861 tcactaccag ctaaatcaac tagataaagt tttccactca gcttttgttc cgtttgtgtg
     1921 ttctcttgtt tgacattaat aagaaatata ctgtgactcc tagagctatg ttcattcata
     1981 tttgtaactg ctacatgtct gttggatttt ccttcatcta tggtatccat aacttcatct
     2041 ggactacata caaaacgctc tgtgcacccc tttacatagg gaactcggtt tttgtcttca
     2101 tgaactgaaa ggttggtctt tgaaacatct aacaggtccc ttatcttatc caaatatatt
     2161 tcaaaatatg aaaccttaat atgaaattcc aaattttcat ccatggagta aatataatta
     2221 aaaatatctt gcactattct tggaataatt cccatgcctt ctggatcatg aagtttaccc
     2281 tccattgtgt gtgtcttccc agaggatgtt tggccgtatg caaatattgt tccattatat
     2341 ccttcaagta catctttaac aatcttcttt gcacagtcat tatacacttg ctcttgagat
     2401 gtgcttgact ggaacacccg atcaaatgca taaggcttgg acgcgatcac gaccgtgtct
     2461 tctccctgaa acttggcgat gtacttgtcg ccgcggttca cttcagactc gttgagaggt
     2521 ctgaagcgac acatcacttt gatgttgcac tcggccaggt ccgccatatg tatatctcct
     2581 tcttaaagtt aaacaaaatt atttctagag ggaaaccgtt gtggtctccc tatagtgagt
     2641 cgtattaatt tcgcgggatc gagatctcgg gcagcgttgg gtcctggcca cgggtgcgca
     2701 tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc
     2761 agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga
     2821 cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga
     2881 agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct
     2941 gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct
     3001 ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat
     3061 gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc
     3121 ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc
     3181 ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc
     3241 tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt
     3301 accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     3361 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     3421 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     3481 gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccattg
     3541 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
     3601 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3661 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3721 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3781 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3841 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3901 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3961 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4021 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4081 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4141 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4201 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4261 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4321 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4381 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4441 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4501 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4561 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4621 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4681 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4741 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4801 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg
     4861 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4921 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4981 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5041 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5101 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     5161 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5221 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5281 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5341 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5401 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5461 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5521 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     5581 aggccctttc gtcttcaaga a
//
