LOCUS       sequence-394929-        8072 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human DNA polymerase iota 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 8072)
  AUTHORS   Ashton NW, Jaiswal N, Cestari Moreno N, Semenova IV, D'Orlando DA, 
            Teatin Latancia M, McIntyre J, Woodgate R, Bezsonova I
  TITLE     A novel interaction between RAD23A/B and Y-family DNA polymerases.
  JOURNAL   J Mol Biol. 2023 Nov 5:168353. doi: 10.1016/j.jmb.2023.168353.
  PUBMED    37935254
REFERENCE   2  (bases 1 to 8072)
  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.
            2023 Nov 5:168353. doi: 10.1016/j.jmb.2023.168353."
COMMENT     Sequence Label: pCMV6-AN-HA_WT POLI
FEATURES             Location/Qualifiers
     source          1..8072
                     /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(3277..3293)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3304..3926
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3503..3522)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3950..3970)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3955..4284
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4135..4270
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4351..5145
                     /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(4405..4424)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5015..5034
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5319..5440
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5356..5375)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5410..5429
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5489..5505)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5502..5524)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5513..5529
                     /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(5537..5543,5544..5561,5562..5567))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5582..5603
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5720..5737)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5891..6479)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5971..5990)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6650..7441,7442..7510))
                     /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     7273..7292
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7511..7615)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7642..8070
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7729..7748)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7939..7960
                     /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 gagctggccg
     1081 acgtgggagc cgctgccagc tcccaggggg tgcacgatca ggtgctgcca actcccaacg
     1141 cttctagtag agtgatcgtg catgtggacc tggattgctt ttacgcccag gtggagatga
     1201 ttagcaatcc tgaactgaag gacaaaccac tgggcgtgca gcagaaatac ctggtggtga
     1261 catgcaacta tgaagctcgc aagctggggg tgaagaaact gatgaatgtg cgggacgcca
     1321 aggagaaatg tccccagctg gtgctggtga acggggagga tctgacccgg tacagagaaa
     1381 tgtcttataa ggtgacagag ctgctggagg agttcagtcc tgtggtggag cgcctgggct
     1441 tcgacgaaaa ttttgtggat ctgacagaga tggtggaaaa gcggctgcag cagctgcagt
     1501 ctgacgagct gagtgccgtg actgtgagtg gacacgtgta caacaatcag tcaattaacc
     1561 tgctggatgt gctgcatatc agactgctgg tgggcagcca gattgctgcc gagatgaggg
     1621 aagctatgta taaccagctg ggactgaccg gatgtgctgg agtggcctca aataagctgc
     1681 tggccaaact ggtcagcggc gtgtttaaac ccaaccagca gacagtgctg ctgcctgagt
     1741 cttgccagca cctgatccat agtctgaatc acatcaagga aattcccggc atcgggtaca
     1801 agactgctaa atgtctggag gccctgggaa tcaacagcgt gcgcgacctg cagacctttt
     1861 cccctaagat tctggagaaa gaactgggca tttccgtggc ccagagaatc cagaagctgt
     1921 ctttcggaga ggacaattcc ccagtgatcc tgtctggccc ccctcagagc ttctccgagg
     1981 aagatagctt taagaaatgc tcaagcgagg tggaagctaa gaacaaaatt gaggaactgc
     2041 tggcctccct gctgaatagg gtgtgtcagg atgggcgcaa acctcacacc gtgcggctga
     2101 tcattcggag atactcctct gagaagcact atggaaggga atcacgccag tgccctattc
     2161 caagccatgt gatccagaag ctgggaacag gcaactatga cgtgatgact cctatggtgg
     2221 acatcctgat gaaactgttc cgcaacatgg tgaatgtgaa gatgcctttc catctgaccc
     2281 tgctgtctgt gtgcttttgt aacctgaaag ctctgaatac agccaagaaa gggctgattg
     2341 actactatct gatgccatct ctgagtacca catcacggag cggaaagcac tccttcaaga
     2401 tgaaagacac tcacatggaa gactttccta aggataaaga gaccaataga gatttcctgc
     2461 caagtggcag gatcgagtca acccggacaa gagaatctcc cctggacact accaacttca
     2521 gtaaggagaa ggatattaac gaatttccac tgtgcagcct gcctgaggga gtggaccagg
     2581 aagtgtttaa gcagctgcct gtggatattc aggaggaaat cctgagtggg aagtcaagag
     2641 agaaattcca ggggaagggc agcgtgtctt gtccactgca cgcttccagg ggcgtgctga
     2701 gcttcttttc caagaaacag atgcaggaca tccccattaa ccctagagat catctgagtt
     2761 caagcaaaca ggtgtcctct gtgtccccat gtgagcccgg cacctctggg tttaacagtt
     2821 caagctcctc ttacatgagt tcacagaagg actattccta ctatctggac aatcggctga
     2881 aagatgaaag aatctctcag ggacctaagg agccacaggg cttccacttt acaaacagca
     2941 atccagctgt gtcagccttc catagctttc ccaacctgca gagcgagcag ctgttctcca
     3001 gaaatcacac aactgacagt cataaacaga ctgtggccac cgattcacac gaaggcctga
     3061 cagagaatag ggaaccagac agcgtggatg agaagattac tttcccatcc gacatcgatc
     3121 cccaggtgtt ttacgagctg cccgaagctg tgcagaaaga gctgctggcc gaatggaaga
     3181 ggaccgggtc cgatttccac atcggacata agtaaggatc caagcttctc gagacgcgtt
     3241 aagcggccgc actcgaggtt taaacggccg gccgcggtca tagctgtttc ctgaacagat
     3301 cccgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact
     3361 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg
     3421 tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag
     3481 acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct
     3541 tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag
     3601 ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga
     3661 cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca
     3721 ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt
     3781 ctgattttaa aataactata ccagcaggag gacgtccaga cacagcatag gctacctggc
     3841 catgcccaac cggtgggaca tttgagttgc ttgcttggca ctgtcctctc atgcgttggg
     3901 tccactcagt agatgcctgt tgaattgggt acgcggccag cttggctgtg gaatgtgtgt
     3961 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4021 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4081 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4141 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4201 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4261 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4321 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4381 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4441 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4501 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4561 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4621 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4681 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4741 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4801 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4861 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4921 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4981 cggctgggtg tggccgaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5041 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5101 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5161 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5221 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5281 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5341 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5401 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5461 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5521 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5581 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5641 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5701 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5761 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5821 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5881 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5941 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6001 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6061 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6121 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6181 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6241 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6301 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6361 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6421 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6481 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6541 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6601 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6661 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6721 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6781 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6841 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6901 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6961 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7021 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7081 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7141 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7201 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7261 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7321 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7381 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7441 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7501 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7561 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7621 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     7681 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     7741 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     7801 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     7861 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     7921 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     7981 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     8041 agctgattta acaaaaattt aacgcgaatt tt
//
