LOCUS       sequence-394904-        8096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the catalytic domain (aa 1-420) of human DNA polymerase 
            iota with a HA tag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8096)
  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 8096)
  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_aa 1-420 POLI
FEATURES             Location/Qualifiers
     source          1..8096
                     /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(3301..3317)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3328..3950
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3527..3546)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3974..3994)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3979..4308
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4159..4294
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4375..5169
                     /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(4429..4448)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5039..5058
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5343..5464
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5380..5399)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5434..5453
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5513..5529)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5526..5548)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5537..5553
                     /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(5561..5567,5568..5585,5586..5591))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5606..5627
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5744..5761)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5915..6503)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5995..6014)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6674..7465,7466..7534))
                     /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     7297..7316
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7535..7639)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7666..8094
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7753..7772)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7963..7984
                     /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 agccaagtaa tctcgagtgg
     2341 atatccatat gaaagggctg attgactact atctgatgcc atctctgagt accacatcac
     2401 ggagcggaaa gcactccttc aagatgaaag acactcacat ggaagacttt cctaaggata
     2461 aagagaccaa tagagatttc ctgccaagtg gcaggatcga gtcaacccgg acaagagaat
     2521 ctcccctgga cactaccaac ttcagtaagg agaaggatat taacgaattt ccactgtgca
     2581 gcctgcctga gggagtggac caggaagtgt ttaagcagct gcctgtggat attcaggagg
     2641 aaatcctgag tgggaagtca agagagaaat tccaggggaa gggcagcgtg tcttgtccac
     2701 tgcacgcttc caggggcgtg ctgagcttct tttccaagaa acagatgcag gacatcccca
     2761 ttaaccctag agatcatctg agttcaagca aacaggtgtc ctctgtgtcc ccatgtgagc
     2821 ccggcacctc tgggtttaac agttcaagct cctcttacat gagttcacag aaggactatt
     2881 cctactatct ggacaatcgg ctgaaagatg aaagaatctc tcagggacct aaggagccac
     2941 agggcttcca ctttacaaac agcaatccag ctgtgtcagc cttccatagc tttcccaacc
     3001 tgcagagcga gcagctgttc tccagaaatc acacaactga cagtcataaa cagactgtgg
     3061 ccaccgattc acacgaaggc ctgacagaga atagggaacc agacagcgtg gatgagaaga
     3121 ttactttccc atccgacatc gatccccagg tgttttacga gctgcccgaa gctgtgcaga
     3181 aagagctgct ggccgaatgg aagaggaccg ggtccgattt ccacatcgga cataagtaag
     3241 gatccaagct tctcgagacg cgttaagcgg ccgcactcga ggtttaaacg gccggccgcg
     3301 gtcatagctg tttcctgaac agatcccggg tggcatccct gtgacccctc cccagtgcct
     3361 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     3421 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     3481 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     3541 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     3601 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     3661 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     3721 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     3781 accactgctc ccttccctgt ccttctgatt ttaaaataac tataccagca ggaggacgtc
     3841 cagacacagc ataggctacc tggccatgcc caaccggtgg gacatttgag ttgcttgctt
     3901 ggcactgtcc tctcatgcgt tgggtccact cagtagatgc ctgttgaatt gggtacgcgg
     3961 ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4021 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4081 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4141 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4201 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4261 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4321 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4381 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4441 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4501 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4561 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4621 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4681 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4741 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4801 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4861 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4921 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4981 ttttctggat tcatcgactg tggccggctg ggtgtggccg accgctatca ggacatagcg
     5041 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5101 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5161 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5221 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5281 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5341 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5401 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5461 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5521 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5581 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5641 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5701 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5761 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5821 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5881 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5941 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6001 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6061 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6121 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6181 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6241 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6301 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6361 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6421 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6481 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6541 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6601 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6661 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6721 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6781 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6841 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6901 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6961 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7021 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7081 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7141 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7201 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7261 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7321 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7381 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7441 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7501 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7561 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7621 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg
     7681 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     7741 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     7801 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     7861 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     7921 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     7981 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     8041 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aatttt
//
