LOCUS       sequence-120027-        5453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  enables eukaryotic expression of N-terminally RGS-6xHis–tagged 
            fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5453)
  AUTHORS   Horn M, Geisen C, Cermak L, Becker B, Nakamura S, Klein C, Pagano M,
            Antebi A
  TITLE     DRE-1/FBXO11-Dependent Degradation of BLMP-1/BLIMP-1 Governs C. 
            elegans Developmental Timing and Maturation.
  JOURNAL   Dev Cell. 2014 Mar 4. pii: S1534-5807(14)00069-0. doi: 
            10.1016/j.devcel.2014.01.028.
  PUBMED    24613396
REFERENCE   2  (bases 1 to 5453)
  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"; journalName: "Dev Cell. 
            2014 Mar 4. pii: S1534-5807(14)00069-0. doi: 
            10.1016/j.devcel.2014.01.028."
COMMENT     Sequence Label: RGS-6xHis-pcDNA3.1-
FEATURES             Location/Qualifiers
     source          1..5453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             917..934
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1047..1064)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1053..1277
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1323..1751
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1410..1429)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1620..1641
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1760..1780)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1765..2094
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1945..2080
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2007..2026
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2161..2955
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2215..2234)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2825..2844
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3129..3250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3166..3185)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3220..3239
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3299..3315)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3299..3315)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3312..3334)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3323..3339
                     /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(3347..3353,3354..3371,3372..3377))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3392..3413
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3530..3547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3701..4286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3781..3800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4457..5248,5249..5317))
                     /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     5080..5099
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5318..5422)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 caccatgaga ggatcgcatc accatcacca tcacgggccc tctagactcg agcggccgcc
      961 actgtgctgg atatctgcag aattccacca cactggacta gtggatccga gctcggtacc
     1021 aagcttaagt ttaaaccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     1081 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     1141 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     1201 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     1261 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     1321 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     1381 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     1441 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     1501 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     1561 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     1621 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     1681 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     1741 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     1801 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     1861 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     1921 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     1981 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     2041 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     2101 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     2161 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     2221 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     2281 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     2341 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     2401 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     2461 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     2521 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     2581 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     2641 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     2701 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     2761 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     2821 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     2881 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     2941 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     3001 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     3061 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     3121 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3181 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3241 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     3301 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     3361 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     3421 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3481 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3541 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3601 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3661 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3721 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3781 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3841 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3901 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3961 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4021 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4081 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4141 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4201 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     4261 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4321 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4381 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4441 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4501 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4561 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4621 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4681 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4741 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4801 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4861 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4921 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4981 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5041 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5101 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5161 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5221 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5281 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5341 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5401 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
