LOCUS       sequence-394925-        8537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human DNA polymerase kappa 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 8537)
  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 8537)
  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 POLK
FEATURES             Location/Qualifiers
     source          1..8537
                     /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"
     RBS             1601..1609
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(3742..3758)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3769..4391
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3968..3987)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4415..4435)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4420..4749
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4600..4735
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4816..5610
                     /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(4870..4889)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5480..5499
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5784..5905
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5821..5840)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5875..5894
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5954..5970)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5967..5989)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5978..5994
                     /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(6002..6008,6009..6026,6027..6032))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6047..6068
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6185..6202)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6356..6944)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6436..6455)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7115..7906,7907..7975))
                     /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     7738..7757
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7976..8080)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8107..8535
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8194..8213)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8404..8425
                     /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 gacagcacca
     1081 aggagaagtg cgactcttat aaggatgacc tgctccttcg gatgggcctg aacgacaata
     1141 aggctggcat ggaaggcctg gacaaggaga agatcaacaa gatcatcatg gaagctacaa
     1201 agggctctag attctacggc aacgagctga agaaagagaa gcaggtgaac cagcggatcg
     1261 agaacatgat gcagcagaag gcccagatca caagccagca gctgagaaag gctcagctgc
     1321 aggtggacag attcgctatg gagctggaac aaagcagaaa cttgagcaac accatcgtgc
     1381 atatcgacat ggacgccttc tacgccgccg ttgagatgcg ggacaacccc gaactgaagg
     1441 acaagcccat cgctgtgggc agcatgagca tgctgtctac tagcaactac cacgccagaa
     1501 gattcggcgt gcgggccgct atgcctggtt tcatcgccaa gagactgtgc cctcagctga
     1561 tcatagttcc acctaacttt gataagtaca gagccgtgtc taaggaggtg aaggaaatcc
     1621 tggctgatta cgaccctaac ttcatggcca tgtctctgga cgaggcctac ctgaacatca
     1681 ccaagcacct ggaggaaagg cagaattggc ccgaggacaa gcgcagatac ttcatcaaga
     1741 tgggcagcag tgtcgagaat gacaatcccg gcaaggaggt gaacaagctg agcgaacacg
     1801 agagatctat cagccctctg ctctttgagg aaagccccag cgacgttcag cctcctggag
     1861 atcctttcca ggtcaatttt gaggagcaga acaacccaca gatcctgcaa aacagcgtgg
     1921 tgttcggcac cagcgcccag gaggtcgtga aggagatccg ctttcggatc gagcagaaga
     1981 caacactgac agccagcgcc ggcatcgccc ctaataccat gctggccaag gtgtgttctg
     2041 acaaaaacaa acctaacggc cagtaccaga tcctgcctaa ccggcaggcc gtgatggact
     2101 tcatcaagga tctgcctatc agaaaagtgt ctggcattgg aaaggtgacc gaaaagatgc
     2161 tgaaggctct gggcatcatc acctgcaccg agctctacca gcaaagagcc ctgctgtccc
     2221 tgctgtttag cgagacaagc tggcactact tcctgcacat cagcctgggc ctgggaagca
     2281 cccacctgac cagagatgga gaaaggaaga gcatgagcgt ggaaagaacc ttcagcgaaa
     2341 tcaacaaggc cgaggaacaa tacagccttt gccaagagct gtgctctgag ctggcccaag
     2401 atctgcagaa agagagactg aaaggcagaa ccgtgacaat caaactgaag aatgtgaatt
     2461 tcgaggtgaa gactagagcc tctacagtga gcagcgtggt gtccaccgcc gaggagattt
     2521 tcgccatcgc caaggagctg ctgaagaccg agatcgacgc cgatttccct caccctctgc
     2581 ggttgagact gatgggagtt agaatcagct ctttccctaa cgaggaggac cggaagcacc
     2641 agcagagaag catcatcggc ttcctgcagg ccggcaacca ggctctaagc gccacagaat
     2701 gtaccctgga aaaaaccgac aaggacaagt tcgtgaaacc cctggagatg agccacaaga
     2761 agtccttttt cgacaagaag cggtccgagc ggaagtggag ccaccaggac acattcaagt
     2821 gcgaggccgt gaacaaacag agctttcaga ccagccagcc attccaggtg ctgaaaaaaa
     2881 agatgaacga aaacctggaa atctccgaga atagcgacga ctgccagatc ctgacatgcc
     2941 ccgtgtgctt cagagctcag ggatgtatct cactggaagc ccttaacaag cacgtggatg
     3001 agtgtctgga cggcccttct atctcagaga actttaagat gttcagctgc agccacgtga
     3061 gcgccacgaa agtgaacaag aaggaaaacg tgcccgccag cagcctgtgt gaaaaacagg
     3121 actacgaggc ccaccctaag atcaaggaga tctctagcgt ggattgcatc gccctggtgg
     3181 ataccatcga caactctagc aaggctgaat ccatcgatgc cctgagcaat aaacattcta
     3241 aggaagaatg cagcagcctg ccgagtaaaa gctttaacat cgagcactgt catcagaaca
     3301 gctccagcac cgtgtcccta gaaaatgaag atgtgggcag cttccggcag gaataccggc
     3361 agccatacct gtgcgaggtg aagaccggcc aagccctggt gtgccctgtg tgcaacgtcg
     3421 aacaaaaaac ctccgatctg acactgttca acgtgcacgt ggacgtgtgc ctgaacaaga
     3481 gctttatcca ggagctgcgg aaggacaagt tcaaccccgt gaaccagcct aaggaatcta
     3541 gcagaagcac cggctcctca tccggcgtgc agaaggccgt gaccagaaca aagagacctg
     3601 gcctgatgac caagtatagc acaagcaaga agattaagcc taacaatccc aagcacaccc
     3661 tggatatctt cttcaagtaa ggatccaagc ttctcgagac gcgttaagcg gccgcactcg
     3721 aggtttaaac ggccggccgc ggtcatagct gtttcctgaa cagatcccgg gtggcatccc
     3781 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     3841 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     3901 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     3961 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     4021 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
     4081 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
     4141 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
     4201 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttaaaataa
     4261 ctataccagc aggaggacgt ccagacacag cataggctac ctggccatgc ccaaccggtg
     4321 ggacatttga gttgcttgct tggcactgtc ctctcatgcg ttgggtccac tcagtagatg
     4381 cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt agggtgtgga
     4441 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4501 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4561 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4621 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4681 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4741 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     4801 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4861 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4921 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4981 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5041 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5101 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5161 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5221 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5281 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5341 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5401 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcc
     5461 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5521 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5581 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5641 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5701 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5761 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     5821 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5881 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5941 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6001 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6061 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6121 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6181 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6241 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6301 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6361 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6421 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6481 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6541 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6601 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6661 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6721 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6781 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6841 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6901 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6961 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7021 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     7081 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7141 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7201 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7261 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7321 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7381 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7441 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7501 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7561 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7621 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7681 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7741 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7801 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7861 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7921 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7981 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     8041 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     8101 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     8161 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     8221 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     8281 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     8341 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     8401 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     8461 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     8521 aatttaacgc gaatttt
//
