LOCUS       sequence-468724-        5511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of mouse HRAS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5511)
  AUTHORS   Fritsch R, de Krijger I, Fritsch K, George R, Reason B, Kumar MS, 
            Diefenbacher M, Stamp G, Downward J
  TITLE     RAS and RHO families of GTPases directly regulate distinct 
            phosphoinositide 3-kinase isoforms.
  JOURNAL   Cell. 2013 May 23;153(5):1050-63. doi: 10.1016/j.cell.2013.04.031.
  PUBMED    23706742
REFERENCE   2  (bases 1 to 5511)
  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"; doi: 
            "10.1016/j.cell.2013.04"; journalName: "Cell"; date: "2013-05-23- 
            23"; volume: "153"; issue: "5"; pages: "1050-63"
COMMENT     Sequence Label: HRAS
FEATURES             Location/Qualifiers
     source          1..5511
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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)."
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..935
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1561..1583)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1728..1746)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1814..1918
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1919..1987,1988..2779)
                     /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     complement(2137..2156)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2950..3538
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3439..3458
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3692..3709
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3782..3859
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3860..4942
                     /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"
     primer_bind     complement(3879..3898)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4991..4996,4997..5014,5015..5021)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5029..5045
                     /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     5053..5069
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5053..5069
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5065..5511,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5085..5102)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5085..5101)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5094..5116)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gttccgcgtg gatccacaga atacaagctt gtggtggtgg
      961 gcgctggagg cgtgggaaag agtgccctga ccatccagct gatccagaac cactttgtgg
     1021 acgagtatga tcccactata gaggactcct accggaaaca ggtggtcatt gatggggaga
     1081 catgtctact ggacatctta gacacagcag gtcaagaaga gtatagtgcc atgcgggacc
     1141 agtacatgcg cacaggggag ggcttcctct gtgtatttgc catcaacaac accaagtcct
     1201 tcgaggacat ccatcagtac agggagcaga tcaagcgggt gaaagattca gatgatgtgc
     1261 caatggtgct ggtgggcaac aagtgtgacc tggctgctcg cactgttgag tctcggcagg
     1321 cccaggacct tgctcgcagc tatggcatcc cctacattga aacatcagcc aagacccggc
     1381 agggcgtgga ggatgccttc tatacactag tccgtgagat tcggcagcat aaattgcgga
     1441 aactgaaccc acccgatgag agtggtcctg gctgcatgag ctgcaaatgt gtgctgtcct
     1501 gagaattcat cgtgactgac tgacgatctg cctcgcgcgt ttcggtgatg acggtgaaaa
     1561 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     1621 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     1681 ccagtcacgt agcgatagcg gagtgtataa ttcttgaaga cgaaagggcc tcgtgatacg
     1741 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     1801 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1861 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     1921 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     1981 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     2041 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     2101 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     2161 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     2221 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     2281 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     2341 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     2401 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     2461 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     2521 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     2581 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     2641 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2701 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     2761 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     2821 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     2881 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2941 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3001 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3061 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     3121 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3181 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3241 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3301 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3361 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3421 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3481 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3541 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     3601 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     3661 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     3721 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcataaattc
     3781 cgacaccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag
     3841 tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg
     3901 tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac
     3961 gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca
     4021 acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca
     4081 cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt
     4141 ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct
     4201 tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat
     4261 tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac
     4321 acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct
     4381 ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc
     4441 gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc
     4501 ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa
     4561 tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat
     4621 gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga
     4681 cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg
     4741 cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa
     4801 gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac
     4861 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     4921 ccgactggaa agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg
     4981 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     5041 aacaatttca cacaggaaac agctatgacc atgattacgg attcactggc cgtcgtttta
     5101 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     5161 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     5221 cgcagcctga atggcgaatg gcgctttgcc tggtttccgg caccagaagc ggtgccggaa
     5281 agctggctgg agtgcgatct tcctgaggcc gatactgtcg tcgtcccctc aaactggcag
     5341 atgcacggtt acgatgcgcc catctacacc aacgtgacct atcccattac ggtcaatccg
     5401 ccgtttgttc ccacggagaa tccgacgggt tgttactcgc tcacatttaa tgttgatgaa
     5461 agctggctac aggaaggcca gacgcgaatt atttttgatg gcgttggaat t
//
