LOCUS       sequence-412138-        6474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ARHGEF2 protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6474)
  TITLE     TREAT-AD plasmids Jun 2023
REFERENCE   2  (bases 1 to 6474)
  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: ARHGEF2A-c052
FEATURES             Location/Qualifiers
     source          1..6474
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     563..582
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(602..679)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     885..904
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     988..1007
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        988..1006
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1007..1031
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             1046..1068
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1079..1096
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1121..1141
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2326..2343
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2396..2414)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2410..2457
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2494..2949
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2581..2600)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2791..2812
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3041..3856)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3041..3856)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3764..3783
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3848..3867
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3978..4566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4467..4486
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4720..4737
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4752..4891
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4817..4836)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4977..4999
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4993..5184)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5993..6474,1..601))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
       61 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
      121 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
      181 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
      241 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
      301 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
      361 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
      421 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
      481 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
      541 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
      601 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
      661 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
      721 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
      781 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
      841 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
      901 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
      961 aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
     1021 ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata tacatatgca
     1081 ccatcatcat catcattctt ctggtgtaga tctgggtacc gagaacctgt acttccaatc
     1141 catggagatg gatgaaaagg actttgcagc ggattcctgg tcattagccg ttgatagctc
     1201 ttttttgcaa cagcacaaaa aggaagttat gaagcaacaa gacgttatct acgaactgat
     1261 ccagaccgag ttacatcatg ttcgtactct gaaaatcatg acccgtttgt ttcgcaccgg
     1321 catgctggag gagcttcacc tggaaccggg tgtcgtccag ggtctgtttc cgtgtgttga
     1381 cgaattgtcc gacatccaca cccgcttcct gtctcaactg ttggagcgtc gccgtcaagc
     1441 cctgtgtccg ggtagcaccc gtaactttgt tattcaccgc ctaggcgacc tgctcattag
     1501 ccagtttagc ggcccgagcg cagaacagat gtgtaaaaca tatagcgagt tctgcagccg
     1561 ccacagcaag gcattgaagc tttacaaaga actttacgcc cgcgacaaac gttttcagca
     1621 gttcattcgt aaggtgaccc gtccggctgt gctcaagcgc cacggcgtgc aggagtgcat
     1681 tctgctggtg acccagcgta ttaccaaata cccgctgctt atctctagaa tcctgcagca
     1741 cagccatggt attgaagagg agcgccaaga cctgacgacc gcgttgggcc tggttaaaga
     1801 actgctgtct aatgtcgacg agggcattta tcagttggaa aagggtgcac gtctgcaaga
     1861 gatttacaat cgtatggatc cgcgagctca gactccggtg ccgggtaaag gcccgttcgg
     1921 tagagaagaa ctcctgcgtc ggaagttgat ccacgatggc tgcctgctgt ggaaaaccgc
     1981 gaccggtcgt ttcaaagacg tcctggttct gcttatgacc gatgtgctgg tgtttctgca
     2041 agagaaggac cagaagtata tatttccgac cctggataaa ccgagcgttg ttagcctgca
     2101 aaacctgatc gtgcgtgaca tcgcgaacca ggagaagggt atgtttctga tttcagccgc
     2161 gcctccggaa atgtatgaag ttcataccgc gagccgtgat gatagaagca cctggattcg
     2221 tgtgatccaa caaagcgtac gcacgtgccc gagccgtgaa taacagtaaa ggtggatacg
     2281 gatccgaatt cgagctccgt cgacaagctt gcggccgcac tcgagcacca ccaccaccac
     2341 cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     2401 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     2461 aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     2521 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2581 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2641 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2701 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2761 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2821 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2881 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     2941 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3001 ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac tcatcgagca
     3061 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     3121 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     3181 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     3241 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     3301 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     3361 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     3421 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     3481 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     3541 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     3601 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     3661 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     3721 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     3781 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc
     3841 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     3901 ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3961 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4021 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4081 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     4141 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4201 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4261 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4321 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4381 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4441 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4501 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4561 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4621 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     4681 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     4741 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4801 aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
     4861 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     4921 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     4981 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     5041 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     5101 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     5161 ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg
     5221 ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca
     5281 tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca
     5341 gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca
     5401 gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt
     5461 ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt
     5521 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg
     5581 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat
     5641 gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
     5701 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
     5761 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
     5821 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
     5881 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     5941 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     6001 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6061 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     6121 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     6181 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     6241 tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact
     6301 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     6361 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     6421 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgcc
//
