LOCUS       sequence-346321-        6652 bp    DNA     circular SYN 15-APR-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 6652)
  TITLE     Gadella lab plasmids
REFERENCE   2  (bases 1 to 6652)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: SspB-iRFP-DHPH1-TRIO
FEATURES             Location/Qualifiers
     source          1..6652
                     /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..1887
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     polyA_signal    3440..3561
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3477..3496)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3531..3550
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3568..4023)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3705..3726)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3917..3936
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4050..4154
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4156..4513
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4156..4347
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4179..4199)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4364..4499
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4548..5342
                     /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             4548..5342
                     /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(4602..4621)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5212..5231
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5530..5549)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5574..5621
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5950..6538
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6439..6458
                     /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 caccatggcg
      961 cgtaaggtcg atctcacctc ctgcgatcgc gagccgatcc acatccccgg cagcattcag
     1021 ccgtgcggct gcctgctagc ctgcgacgcg caggcggtgc ggatcacgcg cattacggaa
     1081 aatgccggcg cgttctttgg acgcgaaact ccgcgggtcg gtgagctact cgccgattac
     1141 ttcggcgaga ccgaagccca tgcgctgcgc aacgcactgg cgcagtcctc cgatccaaag
     1201 cgaccggcgc tgatcttcgg ttggcgcgac ggcctgaccg gccgcacctt cgacatctca
     1261 ctgcatcgcc atgacggtac atcgatcatc gagttcgagc ctgcggcggc cgaacaggcc
     1321 gacaatccgc tgcggctgac gcggcagatc atcgcgcgca ccaaagaact gaagtcgctc
     1381 gaagagatgg ccgcacgggt gccgcgctat ctgcaggcga tgctcggcta tcaccgcgtg
     1441 atgttgtacc gcttcgcgga cgacggctcc gggatggtga tcggcgaggc gaagcgcagc
     1501 gacctcgaga gctttctcgg tcagcacttt ccggcgtcgc tggtcccgca gcaggcgcgg
     1561 ctactgtact tgaagaacgc gatccgcgtg gtctcggatt cgcgcggcat cagcagccgg
     1621 atcgtgcccg agcacgacgc ctccggcgcc gcgctcgatc tgtcgttcgc gcacctgcgc
     1681 agcatctcgc cctgccatct cgaatttctg cggaacatgg gcgtcagcgc ctcgatgtcg
     1741 ctgtcgatca tcattgacgg cacgctatgg ggattgatca tctgtcatca ttacgagccg
     1801 cgtgccgtgc cgatggcgca gcgcgtcgcg gccgaaatgt tcgccgactt cttatcgctg
     1861 cacttcaccg ccgcccacca ccaacgcgtg tacaagtccg gactcagatc tcgagctagg
     1921 aaagagttca taatggctga gctcattcaa actgaaaagg cttatgtaag agacctccgg
     1981 gaatgtatgg atacgtacct gtgggaaatg accagtggcg tggaagagat tccacctggc
     2041 attgtaaaca aagaactcat catcttcgga aacatgcaag aaatctacga atttcataat
     2101 aacatattcc taaaggagct ggaaaaatat gaacagttgc cagaggatgt tggacattgt
     2161 tttgttactt gggcagacaa gtttcagatg tatgtcacat attgcaaaaa taagcctgat
     2221 tctactcagc tgatattgga acatgcaggg tcctattttg acgagataca gcagcgacat
     2281 ggattagcca attccatttc ttcctacctt attaaaccag ttcagcgaat aacgaagtat
     2341 cagctccttt taaaagagct gctgacgtgc tgtgaggaag gaaagggaga gattaaagat
     2401 ggcctggagg tgatgctcag cgtgccgaag cgagccaatg acgccatgca cctcagcatg
     2461 ctggaagggt ttgatgaaaa cattgagtct cagggagaac tcatcctaca ggaatccttc
     2521 caagtgtggg acccaaaaac cttaattcga aagggtcgag aacggcatct cttccttttt
     2581 gaaatgtcct tagtatttag taaagaagtg aaagattcca gtgggagaag caagtacctt
     2641 tataaaagca aattgtttac ctcagagttg ggtgtcacag aacatgttga aggagaccct
     2701 tgcaaatttg cactgtgggt ggggagaaca ccaacttcag ataataaaat tgtccttaag
     2761 gcttccagca tagagaacaa gcaggactgg ataaagcata tccgcgaagt catccaggag
     2821 cggacgatcc acctgaaggg agccctgaag gagcccattc acatccctaa gaccgctccc
     2881 gccacaagac agaagggaag gagggatgga gaggatctgg acagccaagg agacggcagc
     2941 agccagcctg atacgatttc catcgcctca cggacgtctc agaacacgct ggacagcgat
     3001 aagctctctg gtggctgtga gctgacagtg gtgatccatg acttcaccgc ttgcaacagc
     3061 aacgagctga ccatccgacg gggccagacc gtggaagttc tggagcggcc gcatgacaag
     3121 cctgactggt gtctggtgcg gaccactgac cgctccccag cggcagaagg cctggtcccc
     3181 tgtggttcac tgtgcatcgc ccactccaga agtagcatgg aaatggaggg catcttcaac
     3241 cacaaagact cgctctccgt ctccagcaat gacgcctagg aattctgcag tcgacggtac
     3301 cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata ccacatttgt
     3361 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     3421 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     3481 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3541 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     3601 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3661 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3721 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3781 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     3841 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     3901 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     3961 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     4021 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4081 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4141 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     4201 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4261 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4321 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4381 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4441 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4501 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4561 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4621 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4681 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     4741 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4801 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4861 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4921 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4981 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5041 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     5101 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5161 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5221 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5281 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5341 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5401 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5461 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     5521 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     5581 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     5641 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5701 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5761 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     5821 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5881 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5941 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6001 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6061 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6121 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6181 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6241 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6301 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6361 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6421 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6481 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6541 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6601 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
