LOCUS       sequence-154551-        5268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5268)
  TITLE     Bokoch Lab GTPases
REFERENCE   2  (bases 1 to 5268)
  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: pRK5-myc-Rac1-Q61L
FEATURES             Location/Qualifiers
     source          1..5268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        250..629
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        630..833
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     784..804
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     830..854
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1064..1082
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1064..1081
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1095..1114
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1161..1170
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1170..1199
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(1209..1385,1386..1388,1389..1784)
                     /codon_start=1
                     /product="constitutively active mutant of human Rac family 
                     small GTPase 1"
                     /label=Rac1 (Q61L)
                     /translation="QAIKCVVVGDGAVGKTCLLISYTTNAFPGEYIPTVFDNYSANVMV
                     DGKPVNLGLWDTAGLEDYDRLRPLSYPQTDVFLICFSLVSPASFENVRAKWYPEVRHHC
                     PNTPIILVGTKLDLRDDKDTIEKLKEKKLTPITYPQGLAMAKEIGAVKYLECSALTQRG
                     LKTVFDEAIRAVLCPPPVKKRKRKCLLL"
     regulatory      1804..1813
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1817..1951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1854..1873)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1908..1927
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2020..2377
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2020..2211
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2043..2063)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2228..2363
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2397..2414)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2397..2413)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2406..2428)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2626..3081
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2713..2732)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2923..2944
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3098..3117)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3217..3239
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3277..3295)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3363..3467
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3468..3536,3537..4328)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3686..3705)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4499..5087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4988..5007
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5241..5258
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aattaattcg
      241 agctcgcccg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      301 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      361 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      421 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      481 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      541 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      601 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      661 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      721 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      781 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      841 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      901 gggaccgatc cagcctccgc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca
      961 agagtgacgt aagtaccgcc tatagagtct ataggcccac ccccttggct tcgttagaac
     1021 gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg tgacactata
     1081 gaataacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcacct
     1141 cggttctatc gattgaattg gccaccatgg agcagaagct gatctccgag gaggacctgg
     1201 gatccccgca ggccatcaag tgtgtggtgg tgggagacgg agctgtaggt aaaacttgcc
     1261 tactgatcag ttacacaacc aatgcatttc ctggagaata tatccctact gtctttgaca
     1321 attattctgc caatgttatg gtagatggaa aaccggtgaa tctgggctta tgggatacag
     1381 ctggactaga agattatgac agattacgcc ccctatccta tccgcaaaca gatgtgttct
     1441 taatttgctt ttcccttgtg agtcctgcat catttgaaaa tgtccgtgca aagtggtatc
     1501 ctgaggtgcg gcaccactgt cccaacactc ccatcatcct agtgggaact aaacttgatc
     1561 ttagggatga taaagacacg atcgagaaac tgaaggagaa gaagctgact cccatcacct
     1621 atccgcaggg tctagccatg gctaaggaga ttggtgccgt aaaatacctg gagtgctcgg
     1681 cgctcacaca gcgaggcctc aagacagtgt ttgacgaagc gatccgagca gtcctctgcc
     1741 cgcctcccgt gaagaagagg aagagaaaat gcctgctgtt gtaagaattc ctgcagaagc
     1801 ttggccgcca tggcccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     1861 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     1921 actcatcaat gtatcttatc atgtctggat cgggaattaa ttcggcgcag caccatggcc
     1981 tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct
     2041 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2101 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     2161 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     2221 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     2281 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     2341 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctgttaacag cttggcactg
     2401 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     2461 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     2521 tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
     2581 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     2641 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2701 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2761 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2821 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     2881 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2941 ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg attttgccga
     3001 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3061 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     3121 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3181 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3241 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     3301 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     3361 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3421 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3481 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3541 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3601 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3661 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3721 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3781 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3841 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3901 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3961 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4021 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4081 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4141 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4201 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4261 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4321 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4381 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4441 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4501 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4561 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4621 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4681 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4741 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4801 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4861 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4921 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4981 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5041 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5101 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5161 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5221 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
