LOCUS       sequence-373481-        6089 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses GFP-tagged iBox-PAK4cat.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6089)
  AUTHORS   Li TL, Wang Z, You H, Ong Q, Varanasi VJ, Dong M, Lu B, Pasca SP, 
            Cui B
  TITLE     Engineering a Genetically Encoded Magnetic Protein Crystal.
  JOURNAL   Nano Lett. 2019 Oct 9;19(10):6955-6963. doi: 
            10.1021/acs.nanolett.9b02266. Epub 2019 Sep 25.
  PUBMED    31552740
REFERENCE   2  (bases 1 to 6089)
  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: 
            "10.1021/acs.nanolett.9b02266"; journalName: "Nano Lett"; date: 
            "2019-10-9- 9"; volume: "19"; issue: "10"; pages: "6955-6963"
COMMENT     Sequence Label: pXJ40-GFP-iBox-PAK4cat
FEATURES             Location/Qualifiers
     source          1..6089
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..312
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        313..516
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     467..487
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     513..537
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          678..1250
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     1258..1277
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1305..1324
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1305..1323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1336..2049
                     /codon_start=1
                     /gene="mgfp5"
                     /product="GFP with folding enhancement mutations"
                     /label=mgfp5
                     /note="suitable for expression in plants due to removal of 
                     a cryptic intron"
                     /translation="MSKGEELFTGVVPILVELDGDVNGYKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKTRHNIEDGGVQLADHYQQNTPIGDDPVLLPDNHYLSTQSALSKDPNEKRDHMVILE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1366..1394)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     polyA_signal    3182..3303
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3219..3238)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3273..3292
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3337..3354)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3508..4096)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3588..3607)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4267..5058,5059..5127))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4890..4909
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5128..5232)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5300..5318
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5356..5378)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5478..5497
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5564..6019)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5701..5722)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5913..5932
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctagagtccg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
       61 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      121 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      181 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      241 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      301 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      361 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      421 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      481 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
      541 tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc
      601 cgcggccggg aacggtgcat tggaacggac ccctaggctt ttgcaaaaag ctccggatcg
      661 atcctgagaa cttcagggtg agtttgggga cccttgattg ttctttcttt ttcgctattg
      721 taaaattcat gttatatgga gggggcaaag ttttcagggt gttgtttaga atgggaagat
      781 gtcccttgta tcaccatgga ccctcatgat aattttgttt ctttcacttt ctactctgtt
      841 gacaaccatt gtctcctctt attttctttt cattttctgt aactttttcg ttaaacttta
      901 gcttgcattt gtaacgaatt tttaaattca cttttgttta tttgtcagat tgtaagtact
      961 ttctctaatc actttttttt caaggcaatc agggtatatt atattgtact tcagcacagt
     1021 tttagagaac aattgttata attaaatgat aaggtagaat atttctgcat ataaattctg
     1081 gctggcgtgg aaatattctt attggtagaa acaactacat cctggtcatc atcctgcctt
     1141 tctctttatg gttacaatga tatacactgt ttgagatgag gataaaatac tctgagtcca
     1201 aaccgggccc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1261 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attgtaatac gactcactat
     1321 agggcgaatt ccaccatgag taaaggagaa gaacttttca ctggagttgt cccaattctt
     1381 gttgaattag atggtgatgt taatgggtac aaattttctg tcagtggaga gggtgaaggt
     1441 gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt
     1501 ccatggccaa cacttgtcac tactctcact tatggtgttc aatgcttttc aagatatcca
     1561 gatcatatga agcggcacga cttcttcaag agcgccatgc ctgagggata cgtgcaggag
     1621 aggaccatct tcttcaagga cgacgggaac tacaagacac gtgctgaagt caagtttgag
     1681 ggagacaccc tcgtcaacag gatcgagctt aagggaatcg atttcaagga ggacggaaac
     1741 atcctcggcc acaagttgga atacaactac aactcccaca acgtatacat catggccgac
     1801 aagcaaaaga acggcatcaa agccaacttc aagacccgcc acaacatcga agacggcggc
     1861 gtgcaactcg ctgatcatta tcaacaaaat actccaattg gcgatgaccc tgtcctttta
     1921 ccagacaacc attacctgtc cacacaatct gccctttcga aagatcccaa cgaaaagaga
     1981 gaccacatgg tcattcttga gtttgtaacg gctgctggga ttacacatgg catggatgaa
     2041 ctatacaaat caggtggatc cgaagcagag gactggacag cagccctact gaacaggggt
     2101 cgcagtcgcc agcccctggt actaggggac aattgctttg ctgacttggt gcacaactgg
     2161 atggagctgc ctgaggaatt ccccgccgcc cctgctgttc ctgggccccc tggcccccgc
     2221 tcaccacagc gggagccaca gcgagtatcc catgagcagt tccgggctgc cctgcagctg
     2281 gtggtggacc caggcgaccc ccgctcctac ctggacaact tcatcaagat tggcgagggc
     2341 tccacgggca tcgtgtgcat cgccaccgtg cgcagctcgg gcaagctggt ggccgtcaag
     2401 aagatggacc tgcgcaagca gcagaggcgc gagctgctct tcaacgaggt ggtaatcatg
     2461 agggactacc agcacgagaa tgtggtggag atgtacaaca gctacctggt gggggacgag
     2521 ctctgggtgg tcatggagtt cctggaagga ggcgccctca ccgacatcgt cacccacacc
     2581 aggatgaacg aggagcagat cgcagccgtg tgccttgcag tgctgcaggc cctgtcggtg
     2641 ctccacgccc agggcgtcat ccaccgggac atcaagagcg actcgatcct gctgacccat
     2701 gatggcaggg tgaagctgtc agactttggg ttctgcgccc aggtgagcaa ggaagtgccc
     2761 cgaaggaagt cgctggtcgg cacgccctac tggatggccc cagagctcat ctcccgcctt
     2821 ccctacgggc cagaggtaga catctggtcg ctggggataa tggtgattga gatggtggac
     2881 ggagagcccc cctacttcaa cgagccaccc ctcaaagcca tgaagatgat tcgggacaac
     2941 ctgccacccc gactgaagaa cctgcacaag gtgtcgccat ccctgaaggg cttcctggac
     3001 cgcctgctgg tgcgagaccc tgcccagcgg gccacggcag ccgagctgct gaagcaccca
     3061 ttcctggcca aggcagggcc gcctgccagc atcgtgcccc tcatgcgcca gaaccgcacc
     3121 agatgactcg aggcggccgc cccgggctgc aggagctcgg taccagatct tattaaagca
     3181 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3241 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3301 ttatcatgtc tggtcgactc tagactcttc cgcttcctcg ctcactgact cgctgcgctc
     3361 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3421 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3481 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3541 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3601 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3661 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3721 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3781 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3841 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3901 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3961 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4021 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4081 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4141 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4201 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4261 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4321 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4381 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4441 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4501 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4561 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4621 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4681 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4741 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4801 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4861 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4921 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4981 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5041 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5101 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5161 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5221 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     5281 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     5341 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     5401 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     5461 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     5521 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat
     5581 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     5641 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     5701 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     5761 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     5821 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     5881 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     5941 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     6001 gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     6061 cggtgcgggc ctcttcgcta ttacgccag
//
