LOCUS       sequence-346496-        6418 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 6418)
  TITLE     Gadella lab plasmids
REFERENCE   2  (bases 1 to 6418)
  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-mCherry-DHPH1-TRIO
FEATURES             Location/Qualifiers
     source          1..6418
                     /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"
     CDS             955..1662
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1100..1118)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1368..1387
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3206..3327
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3243..3262)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3297..3316
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3334..3789)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3471..3492)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3683..3702
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3816..3920
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3922..4279
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3922..4113
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3945..3965)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4130..4265
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4314..5108
                     /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             4314..5108
                     /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(4368..4387)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4978..4997
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5296..5315)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5340..5387
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5716..6304
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6205..6224
                     /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 caccatggtg
      961 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     1021 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1081 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     1141 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     1201 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     1261 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     1321 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     1381 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     1441 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     1501 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     1561 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     1621 gagggccgcc actccaccgg cggcatggac gagctgtaca agtccggact cagatctcga
     1681 gctaggaaag agttcataat ggctgagctc attcaaactg aaaaggctta tgtaagagac
     1741 ctccgggaat gtatggatac gtacctgtgg gaaatgacca gtggcgtgga agagattcca
     1801 cctggcattg taaacaaaga actcatcatc ttcggaaaca tgcaagaaat ctacgaattt
     1861 cataataaca tattcctaaa ggagctggaa aaatatgaac agttgccaga ggatgttgga
     1921 cattgttttg ttacttgggc agacaagttt cagatgtatg tcacatattg caaaaataag
     1981 cctgattcta ctcagctgat attggaacat gcagggtcct attttgacga gatacagcag
     2041 cgacatggat tagccaattc catttcttcc taccttatta aaccagttca gcgaataacg
     2101 aagtatcagc tccttttaaa agagctgctg acgtgctgtg aggaaggaaa gggagagatt
     2161 aaagatggcc tggaggtgat gctcagcgtg ccgaagcgag ccaatgacgc catgcacctc
     2221 agcatgctgg aagggtttga tgaaaacatt gagtctcagg gagaactcat cctacaggaa
     2281 tccttccaag tgtgggaccc aaaaacctta attcgaaagg gtcgagaacg gcatctcttc
     2341 ctttttgaaa tgtccttagt atttagtaaa gaagtgaaag attccagtgg gagaagcaag
     2401 tacctttata aaagcaaatt gtttacctca gagttgggtg tcacagaaca tgttgaagga
     2461 gacccttgca aatttgcact gtgggtgggg agaacaccaa cttcagataa taaaattgtc
     2521 cttaaggctt ccagcataga gaacaagcag gactggataa agcatatccg cgaagtcatc
     2581 caggagcgga cgatccacct gaagggagcc ctgaaggagc ccattcacat ccctaagacc
     2641 gctcccgcca caagacagaa gggaaggagg gatggagagg atctggacag ccaaggagac
     2701 ggcagcagcc agcctgatac gatttccatc gcctcacgga cgtctcagaa cacgctggac
     2761 agcgataagc tctctggtgg ctgtgagctg acagtggtga tccatgactt caccgcttgc
     2821 aacagcaacg agctgaccat ccgacggggc cagaccgtgg aagttctgga gcggccgcat
     2881 gacaagcctg actggtgtct ggtgcggacc actgaccgct ccccagcggc agaaggcctg
     2941 gtcccctgtg gttcactgtg catcgcccac tccagaagta gcatggaaat ggagggcatc
     3001 ttcaaccaca aagactcgct ctccgtctcc agcaatgacg cctaggaatt ctgcagtcga
     3061 cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca gccataccac
     3121 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     3181 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     3241 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3301 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     3361 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     3421 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     3481 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     3541 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     3601 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3661 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     3721 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3781 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3841 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3901 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     3961 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4021 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4081 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4141 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4201 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4261 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     4321 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4381 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4441 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     4501 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4561 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4621 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4681 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4741 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4801 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     4861 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4921 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4981 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5041 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5101 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5161 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5221 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5281 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     5341 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     5401 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     5461 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     5521 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     5581 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5641 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5701 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5761 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5821 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5881 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5941 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6001 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6061 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6121 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6181 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6241 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6301 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6361 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
