LOCUS       sequence-465430-        5636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5636)
  TITLE     Hurley Lab Plasmids 2025
REFERENCE   2  (bases 1 to 5636)
  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: pCAG/Rab1a(Q70L)-10xHis
FEATURES             Location/Qualifiers
     source          1..5636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     708..727
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(747..824)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(897..914)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1068..1656)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1148..1167)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1827..2618,2619..2687))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2450..2469
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2688..2792)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2860..2878
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     3023..3045
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3136..3165)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             complement(3787..3807)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(3820..4473)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(3840..3862)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     protein_bind    4496..4512
                     /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)."
     promoter        complement(join(4520..4526,4527..4542,4543..4548))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(4650..5177)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     5126..5148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5140..5157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5141..5157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5173..5189)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5173..5189)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    5197..5213
                     /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)."
     promoter        complement(join(5221..5227,5228..5245,5246..5251))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     CDS             complement(join(5300..5636,1..746))
                     /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 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
       61 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
      121 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
      181 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
      241 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
      301 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
      361 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
      421 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
      481 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
      541 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
      601 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
      661 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
      721 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
      781 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtcggaatt tatgcggtgt
      841 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
      901 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
      961 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     1021 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1081 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1141 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1201 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1261 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1321 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1381 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1441 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     1501 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     1561 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     1621 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     1681 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     1741 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     1801 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     1861 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     1921 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     1981 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     2041 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     2101 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca
     2161 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2221 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2281 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2341 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2401 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg
     2461 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     2521 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     2581 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     2641 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     2701 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2761 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     2821 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     2881 tttcgtcttc aagaattata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     2941 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3001 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     3061 ccgaaacgcg cgaggcagat cgtcagtcag tcacgatgcg gccgctcgag tccttatgga
     3121 gttgggatct tattagtgat ggtgatggtg atggtgatgg tgatggcctc cccctgactg
     3181 cttgacgggt gtagactgga ttttgacatt gcttttctcg gcgccgccgg ctgtggcgcc
     3241 agggcccatc cgcttcttga tctcggcggc cattgtcatg aaagactgct ccacatttgt
     3301 ggcgttcttg gcgcttgtct ccaggaaggg gatgcccagg gaatcggcga actccttggc
     3361 tgtggtgtaa tccaccacct tctttgtggt caggtcgcac ttgttgccca ccagcagctt
     3421 attcacgttc tcggaggcat acctgtcgat ctcctgcagc cactgcttca cattgttaaa
     3481 gctctcctga tcggtcacgt cgtacaccac gatgatgccg tgggcgcccc gatagtagga
     3541 gcttgtgatg gttctgaacc gctccaggcc ggctgtgtcc cagatctgca gcttgattgt
     3601 cttgccatcc agctcgatgg tgcggatctt aaagtccacg ccgatggtgc tgatataaga
     3661 ctctgtgtag gtatcgtcgg cgaacctcag cagcagacag ctcttgccca cgccagaatc
     3721 gccaatcagc agcagtttga acaggtaatc atactcaggg ttcatacttg acatggatcc
     3781 catggcgccc tgaaaataca ggttttcggt cgtatcgatt tttggaggat ggtcgccacc
     3841 accaaacgtg gcttgccagc cctgcaaagg ccatgctata tacttgctgg atttcaagta
     3901 cttatcaatt tgtgggatag cttcaatacg ttttttaaaa caaactaatt ttgggaacgc
     3961 atccaggcac attgggtcca tgtataaaac aacatcaaga gcgtcataca acatgaagtc
     4021 aggatgggtt acatgatcac catttaaata tgttttatga cataaacgat cttcgaacat
     4081 tttcagcatt tcaggtagct tgctaagaaa atcaactttg agagtttcaa agtctttact
     4141 atatgcaatt ctcgaaacac cgtatctaat atccaaaacc gctccttcaa gcattgaaat
     4201 ctctgcacgc tcttttggac aaccacccaa catgttgtgc ttgtcagcta tataacgtat
     4261 gatggccata gactgtgtta atttaacatc accatcaata taataaggaa gattgggaaa
     4321 ctccaaaccc aattcaaact ttttgtttcg ccatttatca ccttcatcgc gctcatacaa
     4381 atgctcttca tatttttctt caagatattc caaaagaagt cgagtgggtt gcacaaggcc
     4441 cttaattttc caataaccta gtatagggga catgaatact gtttcctgtg tgaaattgtt
     4501 atccgctcac aattccacac attatacgag ccgatgatta attgtcaaca gctcatttca
     4561 gaatatttgc cagaaccgtt atgatgtcgg cgcaaaaaac attatccaga acgggagtgc
     4621 gccttgagcg acacgaatta tgcagtgatt tacgacctgc acagccatac cacagcttcc
     4681 gatggctgcc tgacgccaga agcattggtg caccgtgcag tcgataagct aattccaacg
     4741 ccatcaaaaa taattcgcgt ctggccttcc tgtagccagc tttcatcaac attaaatgtg
     4801 agcgagtaac aacccgtcgg attctccgtg ggaacaaacg gcggattgac cgtaatggga
     4861 taggtcacgt tggtgtagat gggcgcatcg taaccgtgca tctgccagtt tgaggggacg
     4921 acgacagtat cggcctcagg aagatcgcac tccagccagc tttccggcac cgcttctggt
     4981 gccggaaacc aggcaaagcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     5041 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     5101 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     5161 tccgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5221 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5281 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5341 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     5401 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     5461 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     5521 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     5581 tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgcc
//
