LOCUS       sequence-273239-        5984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length2 HA-tagged RNF126 with an 
            inactivating mutation in the RING domain (C231A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5984)
  AUTHORS   Smith CJ, McGlade CJ
  TITLE     The ubiquitin ligase RNF126 regulates the retrograde sorting of the 
            cation-independent mannose 6-phosphate receptor.
  JOURNAL   Exp Cell Res. 2014 Jan 15;320(2):219-32. doi: 
            10.1016/j.yexcr.2013.11.013. Epub 2013 Nov 23.
  PUBMED    24275455
REFERENCE   2  (bases 1 to 5984)
  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.yexcr.2013.11.013"; journalName: "Exp Cell Res"; date: 
            "2014-01-15- 15"; volume: "320"; issue: "2"; pages: "219-32"
COMMENT     Sequence Label: HA-RNF126 C231A
FEATURES             Location/Qualifiers
     source          1..5984
                     /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             926..952
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             956..982
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1992..2009)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1998..2222
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2268..2696
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2355..2374)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2565..2586
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2705..2725)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2710..3039
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2890..3025
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2952..2971
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3087..3134
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3153..3527
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3657..3778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3694..3713)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3748..3767
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3827..3843)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3827..3843)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3840..3862)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3851..3867
                     /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(3875..3881,3882..3899,3900..3905))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3920..3941
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4058..4075)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4229..4817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4309..4328)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4988..5779,5780..5848))
                     /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     5611..5630
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5849..5953)
                     /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 gtttaaactt aagcttgcca ccatgtaccc atacgatgtt cctgactatg cgggctatcc
      961 ctatgacgtc ccggactatg caggatccga ggcgtcgccg cagcccggac ggtacttctg
     1021 ccactgctgc tcggtagaga tcgtgccgcg cctgccggat tacatctgcc caaggtgcga
     1081 gtctggcttc attgaggagc ttccagaaga gaccaggaac acagagaatg gctcagcccc
     1141 ctccacagcc cccaccgacc agaaccggca gccatttgag aatgtggacc agcacctgtt
     1201 cacgctgcca cagggataca gccagtttgc ctttggcatc ttcgacgata gctttgagat
     1261 tcccacgttc cctcctggag cccaggccga tgatggcagg gaccctgaga gccgacggga
     1321 gagagagcac cagtctcggc atcggtacgg ggcccggcag ccccgtgccc gcctcactgc
     1381 ccgccgggcc actggccggc atgaaggtgt ccctacgctg gaagggatca tccagcagct
     1441 cgtgaatggc atcatctctc cggctgctgt gcccagcctg ggccttggtc cctggggcgt
     1501 cctgcactcg aacccaatgg actacgcctg gggggccaac ggcctggaca ccatcatcac
     1561 gcagctcctc aatcagtttg agaacaccgg ccccccacct gcagacaagg agaagattca
     1621 ggctctcccc acggtcccag tcacagagga acacgtgggc tcagggcttg aggccccagt
     1681 gtgtaaagaa gactatgcac tgggtgagag tgtgcggcag ttgccctgca accacctgtt
     1741 ccacgacagc tgcatcgtgc cctggctgga gcagcatgac agctgcccgg tctgccgtaa
     1801 gagcctcact ggacagaaca cagccaccaa tcccccaggc ttgaccggtg taggtttctc
     1861 ctcgtcctcg tcctcatcct cctccagctc gcccagcaat gagaacgcca caagcaactc
     1921 ctgattctga agatatccag cacagtggcg gccgctcgag tctagagggc ccgtttaaac
     1981 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2041 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2101 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2161 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2221 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     2281 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2341 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2401 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2461 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2521 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2581 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2641 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2701 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2761 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2821 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2881 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2941 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3001 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3061 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     3121 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     3181 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     3241 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     3301 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     3361 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     3421 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     3481 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     3541 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     3601 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     3661 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3721 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3781 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     3841 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3901 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     3961 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4021 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4081 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4141 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4201 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4261 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4321 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4381 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4441 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4501 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4561 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4621 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4681 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4741 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4801 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4861 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4921 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4981 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5041 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5101 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5161 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5221 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5281 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5341 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5401 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5461 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5521 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5581 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5641 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5701 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5761 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5821 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5881 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5941 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
