LOCUS       sequence-394901-        7496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the catalytic domain (aa 321-843) of human DNA repair 
            protein REV1 with a HA tag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7496)
  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 7496)
  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 321-843 REV1
FEATURES             Location/Qualifiers
     source          1..7496
                     /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(2701..2717)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2728..3350
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2927..2946)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3374..3394)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3379..3708
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3559..3694
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3775..4569
                     /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(3829..3848)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4439..4458
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4743..4864
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4780..4799)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4834..4853
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4913..4929)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4926..4948)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4937..4953
                     /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(4961..4967,4968..4985,4986..4991))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5006..5027
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5144..5161)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5315..5903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5395..5414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6074..6865,6866..6934))
                     /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     6697..6716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6935..7039)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7066..7494
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7153..7172)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7363..7384
                     /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 tatccattct agcaccaagt
     1081 ctacatcctc tgtgagcacc ttttccaagg ccgccccttc tgtgccaagc aagccctccg
     1141 attgcaattt catctctaac ttttatagcc actcccggct gcaccacatc agcatgtgga
     1201 agtgtgagct gaccgagttc gtgaatacac tgcagagaca gtctaacggc atctttccag
     1261 gcagggagaa gctgaagaag atgaagaccg gcagaagcgc cctggtggtg accgacacag
     1321 gcgatatgag cgtgctgaac tcccccaggc accagtcttg catcatgcac gtggacatgg
     1381 attgtttctt tgtgagcgtg ggcatcagga acaggccaga cctgaagggc aagccagtgg
     1441 cagtgacctc caatagggga acaggaagag cacctctgag gccaggagca aacccacagc
     1501 tggagtggca gtactatcag aataagatcc tgaagggcaa ggccgccgac atcccagata
     1561 gctccctgtg ggagaatccc gacagcgccc aggccaacgg catcgattct gtgctgagcc
     1621 gcgccgagat cgcctcttgc agctacgagg cccggcagct gggcatcaag aacggcatgt
     1681 tctttggcca cgccaagcag ctgtgcccta atctgcaggc cgtgccctac gacttccacg
     1741 cctacaagga ggtggcccag accctgtatg agacactggc ctcctacacc cacaacatcg
     1801 aggccgtgtc ttgcgacgag gccctggtgg atatcacaga gatcctggcc gagacaaagc
     1861 tgacacctga cgagtttgcc aacgccgtga gaatggagat caaggatcag acaaagtgtg
     1921 ccgcctctgt gggcatcggc agcaatatcc tgctggcaag gatggcaacc cggaaggcaa
     1981 agcctgacgg acagtatcac ctgaagccag aggaggtgga cgatttcatc aggggacagc
     2041 tggtgaccaa cctgccagga gtgggccact ccatggagtc taagctggcc agcctgggca
     2101 tcaagacctg cggcgatctg cagtatatga caatggccaa gctgcagaag gagttcggcc
     2161 ccaagaccgg ccagatgctg tacaggtttt gtcgcggcct ggacgatcgg cctgtgagaa
     2221 cagagaagga gagaaagtcc gtgtctgccg agatcaatta cggcatcagg ttcacccagc
     2281 caaaggaggc cgaggccttt ctgctgagcc tgtccgagga gatccagcgg agactggagg
     2341 ccaccggcat gaagggcaag agactgacac tgaagatcat ggtgaggaag ccaggagcac
     2401 ctgtggagac agccaagttc ggcggccacg gcatctgcga caacatcgca aggaccgtga
     2461 cactggacca ggcaaccgat aatgccaaga tcatcggcaa ggccatgctg aatatgtttc
     2521 acacaatgaa gctgaacatc tccgatatgc ggggcgtggg catccacgtg aaccagctgg
     2581 tgcctaccaa cctgaatcca tctacatgcc caagcagacc ctccgtgcag tctagctaag
     2641 gatccaagct tctcgagacg cgttaagcgg ccgcactcga ggtttaaacg gccggccgcg
     2701 gtcatagctg tttcctgaac agatcccggg tggcatccct gtgacccctc cccagtgcct
     2761 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     2821 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     2881 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     2941 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     3001 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     3061 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     3121 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     3181 accactgctc ccttccctgt ccttctgatt ttaaaataac tataccagca ggaggacgtc
     3241 cagacacagc ataggctacc tggccatgcc caaccggtgg gacatttgag ttgcttgctt
     3301 ggcactgtcc tctcatgcgt tgggtccact cagtagatgc ctgttgaatt gggtacgcgg
     3361 ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3421 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3481 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3541 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3601 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3661 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3721 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3781 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3841 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3901 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3961 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4021 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4081 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4141 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4201 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4261 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4321 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4381 ttttctggat tcatcgactg tggccggctg ggtgtggccg accgctatca ggacatagcg
     4441 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4501 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4561 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4621 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4681 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4741 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4801 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4861 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     4921 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4981 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5041 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5101 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5161 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5221 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5281 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5341 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5401 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5461 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5521 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5581 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5641 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5701 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5761 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5821 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5881 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5941 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6001 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6061 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6121 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6181 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6241 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6301 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6361 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6421 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6481 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6541 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6601 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6661 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6721 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6781 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6841 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6901 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6961 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7021 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg
     7081 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     7141 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     7201 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     7261 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     7321 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     7381 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     7441 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aatttt
//
