LOCUS       sequence-100658-        6084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  30nm SAH scaffold with EF Hand and split GFP attachment sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6084)
  AUTHORS   Norris SR, Soppina V, Dizaji AS, Schimert KI, Sept D, Cai D, 
            Sivaramakrishnan S, Verhey KJ
  TITLE     A method for multiprotein assembly in cells reveals independent 
            action of kinesins in complex.
  JOURNAL   J Cell Biol. 2014 Nov 10;207(3):393-406. doi: 10.1083/jcb.201407086.
            Epub 2014 Nov 3.
  PUBMED    25365993
REFERENCE   2  (bases 1 to 6084)
  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"; doi: 
            "10.1083/jcb.201407086"; journalName: "J Cell Biol"; date: 
            "2014-11-10- 10"; volume: "207"; issue: "3"; pages: "393-406"
COMMENT     Sequence Label: SNAP-EF(N)-30nmSAH-GFP(1-10)
FEATURES             Location/Qualifiers
     source          1..6084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2074..2715
                     /codon_start=1
                     /product="fragment of superfolder GFP consisting of the 
                     first 10 beta-strands"
                     /label=GFP(1-10)
                     /note="associates with GFP11, the 11th beta-strand, to 
                     reconstitute fluorescent GFP (Cabantous et al., 2005; 
                     Kamiyama et al., 2016)"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATIGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     KYKTRAVVKFEGDTLVNRIELKGTDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFTVRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQTVLSKDPNEK"
     polyA_signal    2872..2993
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2909..2928)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2963..2982
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3000..3455)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3137..3158)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3349..3368
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3482..3586
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3588..3945
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3588..3779
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3611..3631)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3796..3931
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3980..4774
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3980..4774
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4034..4053)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4644..4663
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4962..4981)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5006..5053
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5382..5970
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5871..5890
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggacaa ggattgtgaa atgaaacgca ccacactgga cagccctttg
      661 gggaagctgg agctgtctgg ttgtgagcag ggtctgcacg aaataaagct cctgggcaag
      721 gggacgtctg cagctgatgc cgtggaggtc ccagcccccg ctgcggttct cggaggtccg
      781 gagcccctga tgcaatgcac agcctggctg aatgcctatt tccaccagcc cgaggctatc
      841 gaagagttcc ccgtgccggc tcttcaccat cccgttttcc agcaagagtc gttcaccaga
      901 caggtgttat ggaagctgct gaaggttgtg aaattcggag aagtgatttc ttaccagcaa
      961 ttagcagccc tggcaggcaa ccccgcagcc acggcagcag tgggaggagc aatgagaggc
     1021 aatcctgtcc ctatcctgat cccgtgccac agagtggtct gcagcagcgg agccgtgggc
     1081 ggttacgagg gtggactggc cgtgaaggaa tggcttctgg cccatgaagg ccatcggttg
     1141 gggaagccag gcttgggagg gagctcaggt ctggcagggg cctggctcaa gggagcggga
     1201 gctacctcgg gctccccgcc tgctggccga aacgtgtaca agtccggaag cggtggatca
     1261 ggcgctagca tgaaatcccc ggaagaactc aaaggaatct ttgaaaagta cgccgcgaag
     1321 gagggggacc ccaaccaatt gtcgaaggag gagcttaaac tgttgcttca gacggagttc
     1381 ccttcgctgc tcaaggggat gggttcgggg ctcgagaggg atctgggaag cggaggaagc
     1441 ggagaagagg aagaaaagaa acaaaaagaa gaacaagaaa gacttgcaaa agaagaggca
     1501 gagagaaaac aaaaagaaga acaagaaaga cttgcaaaag aagaggcaga gagaaaacaa
     1561 aaggaggaag aagagagaaa acaaaaggaa gaagaagaga gaaaacaaaa ggaggaagaa
     1621 gaaagaaaat taaaggagga acaagaaaga aaagctgcag aagaaaagaa agctaaagaa
     1681 gaagctgaga gaaaggctaa agaagaacaa gaaaggaaag ctgaagaaga gagaaagaag
     1741 aaagaagagg aagaaagact tgaaagagaa agaaaagaga gagaagaaca agaaaagaaa
     1801 gccaaagaag aggcagagag aattgcaaag ttagaggctg aaaagaaggc agaagaagaa
     1861 agaaaagcca aagaagaaga agagagaaaa gccaaagaag aagaggaaag aaagaagaaa
     1921 gaggagcaag aaagacttgc aaaagaaaag gaagaagcag aaagaaaagc tgcagaggaa
     1981 aagaaagcta aagaagaaca agaaagaaaa gaaaaggaag aagcagaaag aaaacaaaga
     2041 ggaagcggag gaagcggaaa gcttggatcc ggaatgtcca aaggagaaga actgtttacc
     2101 ggtgttgtgc caattttggt tgaactcgat ggtgatgtca acggacataa gttctcagtg
     2161 agaggcgaag gagaaggtga cgccaccatt ggaaaattga ctcttaaatt catctgtact
     2221 actggtaaac ttcctgtacc atggccgact ctcgtaacaa cgcttacgta cggagttcag
     2281 tgcttttcga gatacccaga ccatatgaaa agacatgact tttttaagtc ggctatgcct
     2341 gaaggttacg tgcaagaaag aacaatttcg ttcaaagatg atggaaaata taaaactaga
     2401 gcagttgtta aatttgaagg agatactttg gttaaccgca ttgaactgaa aggaacagat
     2461 tttaaagaag atggtaatat tcttggacac aaactcgaat acaattttaa tagtcataac
     2521 gtatacatca ctgctgataa gcaaaagaac ggaattaaag cgaatttcac agtacgccat
     2581 aatgtagaag atggcagtgt tcaacttgcc gaccattacc aacaaaacac ccctattgga
     2641 gacggtccgg tacttcttcc tgataatcac tacctctcaa cacaaacagt cctgagcaaa
     2701 gatccaaatg aaaaaggaac agtcgacggt accgcgggcc cgggatccac cggatctaga
     2761 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2821 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2881 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2941 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     3001 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3061 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3121 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3181 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3241 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3301 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3361 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3421 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     3481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     3601 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3661 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3721 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3781 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3841 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3901 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3961 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4021 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4081 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4141 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4201 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4261 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4321 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4381 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4441 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4501 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4561 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4621 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4681 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4741 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4801 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4861 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4921 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4981 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     5041 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     5101 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     5161 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     5221 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5281 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5341 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5401 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5461 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5521 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5581 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5641 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5701 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5761 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5821 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5881 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5941 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6001 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6061 gataaccgta ttaccgccat gcat
//
