LOCUS       sequence-346296-        6610 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heterodimerization with iLID, Rac1 and RhoG activation .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6610)
  TITLE     Gadella lab plasmids
REFERENCE   2  (bases 1 to 6610)
  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"
COMMENT     Sequence Label: SspB-HaloTag-DHPH1-TRIO
FEATURES             Location/Qualifiers
     source          1..6610
                     /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"
     CDS             604..933
                     /codon_start=1
                     /product="SspB from Haemophilus influenzae, with the 
                     mutations Y11K and A15E to disrupt the natural dimerization
                     of the protein"
                     /label=SspB nano
                     /note="dimerizes with iLID at high affinity to create iLID 
                     nano (Guntas et al., 2015)"
                     /translation="SSPKRPKLLREYYDWLVDNSFTPYLVVDATYLGVNVPVEYVKDGQ
                     IVLNLSASATGNLQLTNDFIQFNARFKGVSRELYIPMGAALAIYARENGDGVMFEPEEI
                     YDELNI"
     regulatory      949..958
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3398..3519
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3435..3454)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3489..3508
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3526..3981)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3663..3684)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3875..3894
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4008..4112
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4114..4471
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4114..4305
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4137..4157)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4322..4457
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4506..5300
                     /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             4506..5300
                     /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(4560..4579)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5170..5189
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5488..5507)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5532..5579
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5908..6496
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6397..6416
                     /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 gctagccacc
      601 atgagctccc cgaaacgccc taagctgctg cgtgaatatt acgattggct ggttgataac
      661 agctttaccc catatctggt ggtggatgcc acatacctgg gcgtgaacgt gcccgtggag
      721 tatgtgaaag acggtcagat cgtgctgaat ctgtctgcaa gtgcgaccgg caacctgcaa
      781 ctgacaaatg attttatcca gttcaacgcc cgctttaagg gcgtgtctcg tgaactgtat
      841 atcccgatgg gtgccgctct ggccatttac gctcgcgaga acggcgatgg tgtgatgttc
      901 gaaccagaag aaatctatga cgagctgaat attggtagtg gaccggtcgc caccatggcc
      961 gagatcggca ccggcttccc cttcgacccc cactacgtgg aggtgctggg cgagcgcatg
     1021 cactacgttg acgtgggccc ccgcgacggc acccccgtgc tgttcctgca cggcaacccc
     1081 accagcagct acgtgtggcg caacatcatc ccccacgtgg cccccaccca ccgcactatc
     1141 gcccccgacc tgatcggcat gggcaagagc gacaagcccg acctgggcta cttcttcgac
     1201 gaccacgtgc gcttcatgga cgccttcatc gaggccctgg gcctggagga ggtggtgctg
     1261 gtgatccacg actggggcag cgccctgggc ttccactggg ccaagcgcaa ccccgagcgc
     1321 gtgaagggca tcgccttcat ggagttcatc cgccccatcc ccacctggga cgagtggccc
     1381 gagttcgccc gcgagacctt ccaggccttc cgcaccaccg acgtgggccg caagctgatc
     1441 atcgaccaga acgtgttcat cgagggcacc ctgcccatgg gcgtggtgcg ccccctgacc
     1501 gaggtggaga tggaccacta ccgcgagccc ttcctgaacc ccgtggaccg cgagcccctg
     1561 tggcgcttcc ccaacgagct gcccatcgcc ggcgagcccg ccaacatcgt ggccctggtg
     1621 gaggagtaca tggactggct gcaccagagc cccgtgccca agctgctgtt ctggggcacc
     1681 cccggcgtgc tgatcccccc cgccgaggcc gcccgcctgg ccaagagcct gcccaacgtt
     1741 aaggccgtgg acatcggccc cggcctgaac ctgctccagg aggacaaccc cgacctgatc
     1801 ggcagcgaga tcgcccgctg gctgagcacc ctggagatca gcggcgtgta caagtccgga
     1861 ctcagatctc gagctaggaa agagttcata atggctgagc tcattcaaac tgaaaaggct
     1921 tatgtaagag acctccggga atgtatggat acgtacctgt gggaaatgac cagtggcgtg
     1981 gaagagattc cacctggcat tgtaaacaaa gaactcatca tcttcggaaa catgcaagaa
     2041 atctacgaat ttcataataa catattccta aaggagctgg aaaaatatga acagttgcca
     2101 gaggatgttg gacattgttt tgttacttgg gcagacaagt ttcagatgta tgtcacatat
     2161 tgcaaaaata agcctgattc tactcagctg atattggaac atgcagggtc ctattttgac
     2221 gagatacagc agcgacatgg attagccaat tccatttctt cctaccttat taaaccagtt
     2281 cagcgaataa cgaagtatca gctcctttta aaagagctgc tgacgtgctg tgaggaagga
     2341 aagggagaga ttaaagatgg cctggaggtg atgctcagcg tgccgaagcg agccaatgac
     2401 gccatgcacc tcagcatgct ggaagggttt gatgaaaaca ttgagtctca gggagaactc
     2461 atcctacagg aatccttcca agtgtgggac ccaaaaacct taattcgaaa gggtcgagaa
     2521 cggcatctct tcctttttga aatgtcctta gtatttagta aagaagtgaa agattccagt
     2581 gggagaagca agtaccttta taaaagcaaa ttgtttacct cagagttggg tgtcacagaa
     2641 catgttgaag gagacccttg caaatttgca ctgtgggtgg ggagaacacc aacttcagat
     2701 aataaaattg tccttaaggc ttccagcata gagaacaagc aggactggat aaagcatatc
     2761 cgcgaagtca tccaggagcg gacgatccac ctgaagggag ccctgaagga gcccattcac
     2821 atccctaaga ccgctcccgc cacaagacag aagggaagga gggatggaga ggatctggac
     2881 agccaaggag acggcagcag ccagcctgat acgatttcca tcgcctcacg gacgtctcag
     2941 aacacgctgg acagcgataa gctctctggt ggctgtgagc tgacagtggt gatccatgac
     3001 ttcaccgctt gcaacagcaa cgagctgacc atccgacggg gccagaccgt ggaagttctg
     3061 gagcggccgc atgacaagcc tgactggtgt ctggtgcgga ccactgaccg ctccccagcg
     3121 gcagaaggcc tggtcccctg tggttcactg tgcatcgccc actccagaag tagcatggaa
     3181 atggagggca tcttcaacca caaagactcg ctctccgtct ccagcaatga cgcctaggaa
     3241 ttctgcagtc gacggtaccg cgggcccggg atccaccgga tctagataac tgatcataat
     3301 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3361 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3421 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3481 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     3541 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3601 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3661 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3721 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3781 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3841 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3901 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3961 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4021 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4081 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     4141 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4201 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4261 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4321 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4381 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4441 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     4501 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4561 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4621 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4681 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4741 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4801 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4861 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4921 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4981 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     5041 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5101 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5161 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5221 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5281 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5341 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5401 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5461 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     5521 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     5581 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     5641 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     5701 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     5761 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5821 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5881 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5941 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6001 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6061 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6121 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6181 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6241 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6301 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6361 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6421 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6481 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6541 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6601 cgccatgcat
//
