LOCUS       sequence-394899-        8066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of WT DNA polymerase eta with an N-terminal HA tag in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8066)
  AUTHORS   McIntyre J, Vidal AE, McLenigan MP, Bomar MG, Curti E, McDonald JP, 
            Plosky BS, Ohashi E, Woodgate R
  TITLE     Ubiquitin mediates the physical and functional interaction between 
            human DNA polymerases eta and iota.
  JOURNAL   Nucleic Acids Res. 2013 Feb 1;41(3):1649-60. doi: 
            10.1093/nar/gks1277. Epub 2012 Dec 16.
  PUBMED    23248005
REFERENCE   2  (bases 1 to 8066)
  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: 
            "10.1093/nar/gks1277"; journalName: "Nucleic Acids Res"; date: 
            "2013-02-1- 1"; volume: "41"; issue: "3"; pages: "1649-60"
COMMENT     Sequence Label: pCMV6-AN-HA_WT POLH
FEATURES             Location/Qualifiers
     source          1..8066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     165..182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     877..897
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     952..971
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1023..1049
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3271..3287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3298..3920
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3497..3516)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3944..3964)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3949..4278
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4129..4264
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4345..5139
                     /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(4399..4418)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5009..5028
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5313..5434
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5350..5369)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5404..5423
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5483..5499)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5496..5518)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5507..5523
                     /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(5531..5537,5538..5555,5556..5561))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5576..5597
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5714..5731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5885..6473)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5965..5984)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6644..7435,7436..7504))
                     /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     7267..7286
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7505..7609)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7636..8064
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7723..7742)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7933..7954
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccacca
     1021 tgtacccata cgatgttcca gattacgctg cgatcgccga tatccatatg gctacaggac
     1081 aggatagagt cgtcgcactg gtggatatgg attgtttctt cgtgcaggtg gagcagcggc
     1141 agaaccctca tctgcgcaac aagccatgcg ctgtggtcca gtacaagtcc tggaaaggcg
     1201 ggggaatcat tgcagtgtct tatgaggcca gagctttcgg agtcaccagg agcatgtggg
     1261 ccgacgatgc caagaagctg tgccccgacc tgctgctggc ccaggtgagg gagtcccgcg
     1321 gcaaggcaaa tctgacaaaa taccgagaag ccagtgtgga ggtcatggaa atcatgtcac
     1381 gctttgccgt gatcgagcga gctagtattg acgaggccta cgtggacctg accagcgccg
     1441 tgcaggagcg gctccagaag ctccagggac agccaatctc agccgacctg ctgcctagca
     1501 cctacattga ggggctgcct cagggaccaa ccacagccga ggaaacagtg cagaaggaag
     1561 gaatgcgcaa acagggcctg ttccagtggc tggacagcct ccagatcgat aacctgacat
     1621 cccccgacct ccagctgact gtgggagcag tgatcgtcga ggaaatgaga gccgctattg
     1681 agagggaaac aggcttccag tgcagcgccg ggatttccca caataaggtg ctggcaaaac
     1741 tggcctgtgg cctgaacaag cctaatcggc agactctggt gtcacacggg agcgtcccac
     1801 agctgttttc ccagatgccc atccgaaaga ttcggtctct gggcgggaaa ctgggcgcca
     1861 gtgtgatcga gattctgggg atcgagtaca tgggagaact gactcagttc accgagtctc
     1921 agctccagag tcactttggc gaaaagaacg ggtcttggct gtatgccatg tgcaggggga
     1981 tcgagcatga tcccgtgaag cctcgccagc tgcctaaaac cattggatgt agcaagaatt
     2041 ttccaggcaa aactgcactg gccaccagag agcaggtgca gtggtggctg ctccagctgg
     2101 cccaggaact ggaggaacgc ctgactaagg accgaaacga caatgatcgg gtggctaccc
     2161 agctggtggt cagcatccgg gtccaggggg ataaaagact gagctccctg cggagatgct
     2221 gtgccctgac aaggtacgac gctcacaaga tgtcccatga tgctttcact gtgatcaaaa
     2281 actgcaatac ctccggcatc cagacagagt ggtctccccc tctgaccatg ctgttcctgt
     2341 gcgccacaaa gtttagtgct tcagcaccct ctagttcaac agacattact tcttttctga
     2401 gctccgatcc ttctagtctg ccaaaggtgc ccgtcacttc aagcgaggct aaaacccagg
     2461 gaagcggccc cgctgtgaca gcaactaaga aagcaactac ctccctggaa tctttctttc
     2521 agaaggcagc cgagagacag aaggtcaaag aagccagcct gtcctctctg accgctccca
     2581 cacaggcacc tatgagcaac tccccttcta aaccatctct gcccttccag accagtcagt
     2641 caaccggcac agagcccttc tttaagcaga aaagtctgct gctgaagcag aaacagctga
     2701 acaatagttc agtgagctcc ccacagcaga acccctggtc aaattgcaag gccctgccta
     2761 atagcctgcc aactgagtac cccgggtgcg tgcctgtctg tgaaggagtg agcaagctgg
     2821 aggaatctag taaagccacc cctgctgaga tggacctggc tcacaacagc cagtccatgc
     2881 atgcatcaag cgcctctaag agtgtgctgg aagtcacaca gaaagccact ccaaatccca
     2941 gcctgctggc cgccgaggat caggtgccat gcgaaaagtg tggcagcctg gtgcccgtgt
     3001 gggacatgcc agagcacatg gattatcatt tcgccctgga actccagaag agcttcctcc
     3061 agcctcactc ctctaaccca caggtggtca gtgccgtgtc acatcagggc aagaggaacc
     3121 ctaaatcccc actggcttgt acaaataaga gaccaagacc cgaaggaatg cagaccctgg
     3181 agagcttttt caaaccactg actcactaag gatccaagct tctcgagacg cgttaagcgg
     3241 ccgcactcga ggtttaaacg gccggccgcg gtcatagctg tttcctgaac agatcccggg
     3301 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     3361 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     3421 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     3481 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     3541 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     3601 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     3661 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     3721 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     3781 ttaaaataac tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc
     3841 caaccggtgg gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact
     3901 cagtagatgc ctgttgaatt gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta
     3961 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4021 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4081 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4141 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4201 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4261 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4321 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4381 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4441 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4501 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4561 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4621 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4681 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4741 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4801 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4861 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4921 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4981 ggtgtggccg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5041 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5101 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5161 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5221 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5281 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5341 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5401 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5461 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5521 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5581 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5641 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5701 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5761 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5821 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5881 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5941 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6001 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6061 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6121 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6181 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6241 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6301 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6361 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6421 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6481 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6541 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6601 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6661 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6721 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6781 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6841 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6901 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6961 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7021 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7081 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7141 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7201 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7261 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7321 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7381 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7441 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7501 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7561 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7621 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     7681 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     7741 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     7801 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     7861 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     7921 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     7981 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     8041 tttaacaaaa atttaacgcg aatttt
//
