LOCUS       sequence-175237-        5225 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express POLR2G (RPB7) tagged with VENUS at C terminus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5225)
  AUTHORS   Xu Y, Anderson DE, Ye Y
  TITLE     The HECT domain ubiquitin ligase HUWE1 targets unassembled soluble 
            proteins for degradation.
  JOURNAL   Cell Discov. 2016 Nov 8;2:16040. eCollection 2016.
  PUBMED    27867533
REFERENCE   2  (bases 1 to 5225)
  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: "Cell 
            Discov."; date: "2016-11-8"; volume: "2"; pages: "16040. eCollection
            2016"
COMMENT     Sequence Label: pVENUS-N1-POLR2G-GFP
FEATURES             Location/Qualifiers
     source          1..5225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..600
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     501..520
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        775..1078
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1079..1282
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1233..1253
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1879..1888
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1885..1887,1888..1890,1891..2604)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     ERVTAAGITLGMDELYK"
     primer_bind     complement(1930..1951)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2191..2210)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2727..2848
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2764..2783)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2818..2837
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2855..3310)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2992..3013)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3204..3223
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3337..3441
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3443..3800
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3443..3634
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3466..3486)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3651..3786
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3835..4629
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3835..4629
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3889..3908)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4499..4518
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4817..4836)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4861..4908
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
       61 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      121 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
      181 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      241 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      301 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      361 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      421 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      481 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      541 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      601 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      661 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
      721 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      781 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      841 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      901 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      961 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     1021 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     1081 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     1141 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     1201 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     1261 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac
     1321 tcagatctcg agctcaagct tgccaccatg ttctaccata tctccctaga gcacgaaatc
     1381 ctgctgcacc cgcgctactt cggccccaac ttgctcaaca cggtgaagca gaagctcttc
     1441 accgaggtgg aggggacctg cacagggaag tatggctttg taattgctgt caccaccatt
     1501 gacaatattg gtgctggtgt gatccagcca ggccgaggct ttgtccttta tccagttaag
     1561 tacaaggcca ttgttttccg gccatttaaa ggggaggtcg tggatgctgt tgtcactcag
     1621 gtcaacaagg ttggactctt cacagaaatt gggcccatgt cttgcttcat ctctcgacat
     1681 tccatccctt cagagatgga gtttgatcct aactccaacc caccatgtta caagacaatg
     1741 gatgaggata ttgtgattca gcaggacgat gagatccgct taaagattgt ggggacccgt
     1801 gtggacaaga atgacatttt tgctattggc tccctgatgg acgattactt ggggcttgta
     1861 agccgggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     1921 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1981 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc
     2041 aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcct gcagtgcttc
     2101 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2161 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     2221 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     2281 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     2341 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     2401 gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     2461 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag caaagacccc
     2521 aacgagaagc gcgatcacat ggtcctgctg gaacgcgtga ccgccgccgg gatcactctc
     2581 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
     2641 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2701 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2761 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2821 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     2881 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2941 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3001 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3061 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3121 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3181 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3241 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3301 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3361 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3421 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3481 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3541 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3601 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3661 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3721 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3781 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3841 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3901 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3961 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     4021 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4081 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4141 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4201 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4261 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4321 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     4381 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4441 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4501 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4561 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4621 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4681 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4741 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4801 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     4861 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     4921 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     4981 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     5041 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     5101 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5161 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5221 agatc
//
