LOCUS       sequence-488268-        7414 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Gateway compatible vector expressing PARG (E755/756A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7414)
  AUTHORS   Nie L, Wang C, Huang M, Liu X, Feng X, Tang M, Li S, Hang Q, Teng H,
            Shen X, Ma L, Gan B, Chen J
  TITLE     DePARylation is critical for S phase progression and cell survival.
  JOURNAL   Elife. 2024 Apr 5;12:RP89303. doi: 10.7554/eLife.89303.
  PUBMED    38578205
REFERENCE   2  (bases 1 to 7414)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2024-04-5"; volume: "12:RP89303. doi"; pages: " 
            10.7554/eLife.89303"
COMMENT     Sequence Label: pJFT7_nHalo_DC(r4)_PARG_Mutation
FEATURES             Location/Qualifiers
     source          1..7414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(114..133)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      927..1515
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1416..1435
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1669..1686
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1715..1762)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1758..1776
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    1796..1820
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4754..4778)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(4794..4814)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             complement(4827..5717)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     misc_feature    5722..6183
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     6110..6127
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(6233..6252)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6234..6252)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6259..6276)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6259..6275)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6268..6290)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6416..6871
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6503..6522)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6713..6734
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6940..6959)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7059..7081
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7119..7137)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7205..7309
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7310..7378,7379..7414,1..756)
                     /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"
ORIGIN
        1 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
       61 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
      121 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
      181 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
      241 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
      301 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
      361 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
      421 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
      481 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
      541 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
      601 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
      661 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
      721 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
      781 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
      841 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
      901 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
      961 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     1021 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     1081 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     1141 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     1201 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     1261 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     1321 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     1381 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     1441 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     1501 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     1561 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     1621 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     1681 agcgaggaag cccggatata gttcctcctt tcagcaaaaa acccctcaag acccgtttag
     1741 aggccccaag gggttatgct agttattgct cagcggacta gctagtgttt aaactaccac
     1801 tttgtacaag aaagctgggt caaggtccct gtcctttgcc ctgaatggtc agcggtctct
     1861 gcacaggact cgacagcatg gtatatgaat ggataaagct tgatgtctgg tccaggggtg
     1921 gaacagtttc tgcattcttc attgtagtat cgtagcaaca gcttatacac atctccaaca
     1981 gtgagtttcc tttcagtaag gaaaatgtgc atgctgtaaa tgtctctcat caattctgag
     2041 tccccaaagg tgaaataaac cacatctcgc tcagctgcag cagctgccaa tatctgtatt
     2101 aaggctttta acctggcatc acccccaaag gcaccacagc cccagtttcc tgtggccact
     2161 gcagaaagat tctctgaaga aactccagga cggagaaatc cacagtaagc cttgttcagc
     2221 tcgcgtctca ttttctcagg cacaaactga tcgaggtagc gtctgaagtg aagagcatcg
     2281 atggcaacga tctcagtgca gcgccgctgc cagtcgtccc tttcactccc atcttcgtgg
     2341 ctccgggacc aacgatatgt ctcagcatag cctgtgtatt cactgtactg ctcagtacct
     2401 gtgataatta gacattcatt gtgatccagc acctcagtga agagccgtga aataatcaac
     2461 tcaggattga ttaaaaagcg gattgctgct tgcacaagtc ctgcactggt tacaccacct
     2521 ccaacaaaac gatttgcaaa atccacctgt agcatgcctt ggccattttc ttctatggta
     2581 ccttcgtaag tgacatgcaa tcgtgtcaag ggtttttcac atctttccca ttctggaaaa
     2641 tcttcaagac tctgtcttgt aaatgtcacc aacccagtag gttttttctc tgtgactctt
     2701 ctaaagtagc agaagagcgt tttaagtttc tccggtttcc ttgatgaacg tccctcaaac
     2761 aatcgattga agttaatgtc tgggtaacta gaatactccg atttcatctt agcatttcgt
     2821 cgtggaaatg tgcagaagaa agcattagct aaaagactgg caatctgttc ctgcgacatt
     2881 gtgatggaat gattcatctt ctgtttcagg agtggtattg gctgggtgca aatatttggc
     2941 agacagagtg caattttcac catatcaggc aagatggact gatataaatg ttgagcttct
     3001 gcttcttcaa gtaccttatc ccagaaatcg atcaaagctg taaagtccca tttcttagaa
     3061 tatgccacat tgtatttcag aatagcatcc ttcaagtttt ggggtcgtgt aaatttgttg
     3121 agaagtgcag tctgaatgag ctcccaccgg ctccccgcag ttcgctcacc attctcatct
     3181 tccactgggt acaaattttg ttctgaacaa ggcattttaa catgcttgtt atcccacaaa
     3241 tctttataat gtgttggaaa aggtttagga acttctcctg ctcgcaaaag atctacccga
     3301 atagttactg tgtgattggc agatggtctc aagagaggca gccggatccc acaccgaggc
     3361 attcttctca tctcctcaat gggagttcca agccacttct tatctggaga aaggtgaggt
     3421 ggaacgtatt tagggatctt cctttctgtt ctttgatgtt tggtttccca ctgttctttt
     3481 cttctgtcct ctgctttggg cagtctcatg aaatgatctg tgattttaga atcctttttt
     3541 ccatgttgct tagaatttct gcattctaca ttcaggctag aaatatttcc aggtagttta
     3601 gcatttaaat cattcattcc agtgcgactc tctcctcctt caaattggaa atgtaatcta
     3661 acttcaccgc ccttagtaga gtaccgtttc ctaaattcaa tgtcagcgtc tcttgcttgg
     3721 aaccttgaag gtttatttgc tgtttgggag gaactaccat cttcttgttc atcaaaacct
     3781 ggacttgtct cctcatctgc ttctgagtct tgacaactat ttttagaatt atccacatcc
     3841 atcggtgact cgggttcact ttccttctca aatggaggag aatttcctag gcaactttct
     3901 tgtctagtca atttgttgtc attttttggc ccagtaccaa catcctcaga gccaacatct
     3961 gacaatggac tctctggcac cacatctatc tcatcttgct gacaacttgc acagtcttcc
     4021 ccaggatcgc aagacttgct gcacttctgg tggcttttgg cttctctggc ctgttcatct
     4081 tctgtagtct gctttgcatt tgcaagcttt acagttgtga gaaactgttg attgtctcta
     4141 ttctcttcac tatctgtgtc actgtgatca tcattttgag gtgaccgatc aatgttagca
     4201 ttactaaact gctctggtac cagggttact gtttgaggtt cactttccaa aagctgctcc
     4261 gtgtgtttcc cttcattttg ccacttacac attgctgcag tctgatgctg gttcaaatac
     4321 tgtgtacttt tttcagtggg tgacttatca agacttagct gagaaacatt ttctaatttt
     4381 tctacattat gttggtaaaa gttatctttt tgtacagaac tcatcatgga ttctattctt
     4441 gtattgttgt tttctttact atccaaactt tctgattccg ctgtcttgat tcctttagtg
     4501 tccatccaac tggtaatagt cttttgtttg aaaacaagcg aggtggcgct gcctctgtgc
     4561 tgtcccgccc gccctgggac gcaggctggc gaggacggtg ggaccctgaa ctgcacgtga
     4621 gcgtccttgg ggtcgaggac gcgcctctgc ctgctgggaa agctccgggc gtccgaagca
     4681 gccggcgaag ttgtagcggc gccccagcgg ggtcgcttgg tgcagggttc acagccgggg
     4741 cccgcattca taaagcctgc ttttttgtac aaacttgtgg cgatcgcgtt atcgctctga
     4801 aagtacagat cctcagtggt tggctcgccg gaaatctcga gcgtcgacag ccagcgcgcg
     4861 atctcgctgc cgatcaggtc cgggttgtct tcttgcagca gattcagacc cgggccgatg
     4921 tccacagcct tgcagttagg caggcttttg gccaggcgag cggcttcggc cggtgggatc
     4981 agaacgcctg gggtgcccca gaacagcagc ttcgggacag gggactggtg cagccagtcc
     5041 atgtattctt cgaccagcgc gacgatgttc gctggctcac cggcgattgg cagctcgttt
     5101 gggaagcgcc acagtggctc gcggtcaaca ggattcagga acggctcgcg gtaatggtcc
     5161 atctcgactt cagtcagcgg gcggacgaca cccatcggca gcgtaccctc gataaaaacg
     5221 ttctgatcga tgatcagctt gcggccgacg tcggtggtgc ggaaggcctg gaaggtctcg
     5281 cgggcaaatt ctggccattc gtcccaggtc gggatagggc ggatgaactc cataaatgca
     5341 atacctttga cgcgctctgg attgcgcttg gcccagtgga aacccagagc ggagccccag
     5401 tcgtgaatga ccaggacgac ctcttccaga cccagggctt cgatgaaggc atccatgaag
     5461 cggacgtggt cgtcgaagaa ataacccagg tctggtttgt cggatttgcc cataccgatc
     5521 aggtctggag caatgcagcg atgggtcggt gcaacatgcg ggatgatgtt gcgccacacg
     5581 taggaggagg tcgggttacc gtgcaggaac agcacagggg tgccatcgcg cggaccaaca
     5641 tcgacgtagt gcatgcgctc gcccaggact tccacataat gggggtcgaa tggaaagcca
     5701 gtaccgattt ctgccatggt attatcatcg tgtttttcaa aggaaaacca cgtccccgtg
     5761 gttcgggggg cctagacgtt ttttaacctc gactaaacac atgtaaagca tgtgcaccga
     5821 ggccccagat cagatcccat acaatggggt accttctggg catccttcag ccccttgttg
     5881 aatacgcttg aggtgagcca tttgactctt tccacaacta tccaactcac aacgtggcac
     5941 tggggttgtg ccgcctttgc aggtgtatct tatacacgtg gcttttggcc gcagaggcac
     6001 ctgtcgccag gtggggggtt ccgctgcctg caaagggtcg ctacagacgt tgtttgtctt
     6061 caagaagctt ccagaggaac tgcttccttc acgacattca acagaccttg cattcctttg
     6121 gcgagagggg aaagacccct aggaatgctc gtcaagaaga cagggccagg tttccgggcc
     6181 ctcacattgc caaaagacgg caatatggtg gaaaataacc ggaattaatt cgccctatag
     6241 tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     6301 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     6361 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
     6421 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     6481 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     6541 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     6601 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     6661 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6721 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6781 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6841 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     6901 tgtgcggtat ttcacaccgc atatatggtg cactctcagt acaatctgct ctgatgccgc
     6961 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     7021 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     7081 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     7141 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     7201 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     7261 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     7321 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     7381 cccagaaacg ctggtgaaag taaaagatgc tgaa
//
