LOCUS       sequence-267280-        8093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of DNA polymerase iota residues 1-420 with an 
            N-terminal FLAG-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8093)
  AUTHORS   Ashton NW, Valles GJ, Jaiswal N, Bezsonova I, Woodgate R
  TITLE     DNA Polymerase iota Interacts with Both the TRAF-like and UBL1-2 
            Domains of USP7.
  JOURNAL   J Mol Biol. 2021 Jan 22;433(2):166733. doi: 
            10.1016/j.jmb.2020.166733. Epub 2020 Dec 3.
  PUBMED    33279577
REFERENCE   2  (bases 1 to 8093)
  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: "J Mol 
            Biol."; date: "2021-01-22"; volume: "433(2):166733. doi"; pages: " 
            10.1016/j.jmb.2020.166733. Epub 2020 Dec 3"
COMMENT     Sequence Label: pCMV6-AN-DDK_1-420 POLI
FEATURES             Location/Qualifiers
     source          1..8093
                     /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"
     regulatory      1014..1023
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1023..1046
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3298..3314)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3325..3947
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3524..3543)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3971..3991)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3976..4305
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4156..4291
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4372..5166
                     /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(4426..4445)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5036..5055
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5340..5461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5377..5396)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5431..5450
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5510..5526)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5523..5545)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5534..5550
                     /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(5558..5564,5565..5582,5583..5588))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5603..5624
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5741..5758)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5912..6500)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5992..6011)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6671..7462,7463..7531))
                     /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     7294..7313
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7532..7636)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7663..8091
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7750..7769)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7960..7981
                     /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 tggattacaa ggatgacgac gataaggcga tcgccgatat ccatatggag ctggccgacg
     1081 tgggagccgc tgccagctcc cagggggtgc acgatcaggt gctgccaact cccaacgctt
     1141 ctagtagagt gatcgtgcat gtggacctgg attgctttta cgcccaggtg gagatgatta
     1201 gcaatcctga actgaaggac aaaccactgg gcgtgcagca gaaatacctg gtggtgacat
     1261 gcaactatga agctcgcaag ctgggggtga agaaactgat gaatgtgcgg gacgccaagg
     1321 agaaatgtcc ccagctggtg ctggtgaacg gggaggatct gacccggtac agagaaatgt
     1381 cttataaggt gacagagctg ctggaggagt tcagtcctgt ggtggagcgc ctgggcttcg
     1441 acgaaaattt tgtggatctg acagagatgg tggaaaagcg gctgcagcag ctgcagtctg
     1501 acgagctgag tgccgtgact gtgagtggac acgtgtacaa caatcagtca attaacctgc
     1561 tggatgtgct gcatatcaga ctgctggtgg gcagccagat tgctgccgag atgagggaag
     1621 ctatgtataa ccagctggga ctgaccggat gtgctggagt ggcctcaaat aagctgctgg
     1681 ccaaactggt cagcggcgtg tttaaaccca accagcagac agtgctgctg cctgagtctt
     1741 gccagcacct gatccatagt ctgaatcaca tcaaggaaat tcccggcatc gggtacaaga
     1801 ctgctaaatg tctggaggcc ctgggaatca acagcgtgcg cgacctgcag accttttccc
     1861 ctaagattct ggagaaagaa ctgggcattt ccgtggccca gagaatccag aagctgtctt
     1921 tcggagagga caattcccca gtgatcctgt ctggcccccc tcagagcttc tccgaggaag
     1981 atagctttaa gaaatgctca agcgaggtgg aagctaagaa caaaattgag gaactgctgg
     2041 cctccctgct gaatagggtg tgtcaggatg ggcgcaaacc tcacaccgtg cggctgatca
     2101 ttcggagata ctcctctgag aagcactatg gaagggaatc acgccagtgc cctattccaa
     2161 gccatgtgat ccagaagctg ggaacaggca actatgacgt gatgactcct atggtggaca
     2221 tcctgatgaa actgttccgc aacatggtga atgtgaagat gcctttccat ctgaccctgc
     2281 tgtctgtgtg cttttgtaac ctgaaagctc tgaatacagc caagtaatct cgagtggata
     2341 tccatatgaa agggctgatt gactactatc tgatgccatc tctgagtacc acatcacgga
     2401 gcggaaagca ctccttcaag atgaaagaca ctcacatgga agactttcct aaggataaag
     2461 agaccaatag agatttcctg ccaagtggca ggatcgagtc aacccggaca agagaatctc
     2521 ccctggacac taccaacttc agtaaggaga aggatattaa cgaatttcca ctgtgcagcc
     2581 tgcctgaggg agtggaccag gaagtgttta agcagctgcc tgtggatatt caggaggaaa
     2641 tcctgagtgg gaagtcaaga gagaaattcc aggggaaggg cagcgtgtct tgtccactgc
     2701 acgcttccag gggcgtgctg agcttctttt ccaagaaaca gatgcaggac atccccatta
     2761 accctagaga tcatctgagt tcaagcaaac aggtgtcctc tgtgtcccca tgtgagcccg
     2821 gcacctctgg gtttaacagt tcaagctcct cttacatgag ttcacagaag gactattcct
     2881 actatctgga caatcggctg aaagatgaaa gaatctctca gggacctaag gagccacagg
     2941 gcttccactt tacaaacagc aatccagctg tgtcagcctt ccatagcttt cccaacctgc
     3001 agagcgagca gctgttctcc agaaatcaca caactgacag tcataaacag actgtggcca
     3061 ccgattcaca cgaaggcctg acagagaata gggaaccaga cagcgtggat gagaagatta
     3121 ctttcccatc cgacatcgat ccccaggtgt tttacgagct gcccgaagct gtgcagaaag
     3181 agctgctggc cgaatggaag aggaccgggt ccgatttcca catcggacat aagtaaggat
     3241 ccaagcttct cgagacgcgt taagcggccg cactcgaggt ttaaacggcc ggccgcggtc
     3301 atagctgttt cctgaacaga tcccgggtgg catccctgtg acccctcccc agtgcctctc
     3361 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     3421 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     3481 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     3541 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     3601 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     3661 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     3721 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     3781 actgctccct tccctgtcct tctgatttta aaataactat accagcagga ggacgtccag
     3841 acacagcata ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc
     3901 actgtcctct catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca
     3961 gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4021 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4081 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4141 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4201 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4261 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4321 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4381 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4441 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4501 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4561 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4621 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4681 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4741 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4801 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4861 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4921 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4981 tctggattca tcgactgtgg ccggctgggt gtggccgacc gctatcagga catagcgttg
     5041 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5101 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5161 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5221 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5281 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5341 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5401 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5461 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5521 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5581 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5641 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5701 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5761 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5821 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5881 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5941 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6001 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6061 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6121 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6181 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6241 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6301 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6361 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6421 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6481 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6541 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6601 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6661 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6721 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6781 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6841 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6901 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6961 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7021 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7081 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7141 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7201 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7261 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7321 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7381 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7441 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7501 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7561 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7621 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     7681 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     7741 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     7801 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     7861 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     7921 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     7981 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     8041 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat ttt
//
