LOCUS       Exported                7275 bp ds-DNA     circular SYN 15-SEP-2021
DEFINITION  Bacterial expression of PARP1 lacking CAT domain.
ACCESSION   .
VERSION     .
KEYWORDS    pET28_6xHis-PARP1delCAT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7275)
  TITLE     HOLD-Conaway Lab Plasmids
REFERENCE   2  (bases 1 to 7275)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 15, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7275
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     960..979
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(999..1076)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1282..1301
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1385..1404
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1385..1403
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1404..1428
                     /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)."
     RBS             1443..1465
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1484..1501
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1511..1528
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             3524..3541
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3594..3612)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3608..3655
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3692..4147
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3779..3798)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3989..4010
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4239..5054)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4962..4981
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5046..5065
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5176..5764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5665..5684
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5918..5935
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5950..6089
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6015..6034)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6175..6197
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6191..6382)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(7191..998)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
       61 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
      121 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
      181 tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
      241 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
      301 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
      361 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
      421 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
      481 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
      541 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
      601 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
      661 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
      721 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
      781 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
      841 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
      901 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
      961 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     1021 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     1081 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     1141 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
     1201 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     1261 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     1321 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg
     1381 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tctagaaata
     1441 attttgttta actttaagaa ggagatatac catgggcagc agccatcacc atcatcatca
     1501 cagcagcggc ctggtgccgc gcggcagcca tatgatggcg gagtcttctg ataagctgta
     1561 tcgagtcgag tacgccaaga gcgggcgcgc ctcttgcaag aaatgcagcg agagcatccc
     1621 caaggactcg ctccggatgg ccatcatggt gcagtcgccc atgtttgatg gaaaagtccc
     1681 acactggtac cacttctcct gcttctggaa ggtgggccac tccatccggc accctgacgt
     1741 tgaggtggat gggttctctg agcttcggtg ggatgaccag cagaaagtca agaagacagc
     1801 ggaagctgga ggagtgacag gcaaaggcca ggatggaatt ggtagcaagg cagagaagac
     1861 tctgggtgac tttgcagcag agtatgccaa gtccaacaga agtacgtgca aggggtgtat
     1921 ggagaagata gaaaagggcc aggtgcgcct gtccaagaag atggtggacc cggagaagcc
     1981 acagctaggc atgattgacc gctggtacca tccaggctgc tttgtcaaga acagggagga
     2041 gctgggtttc cggcccgagt acagtgcgag tcagctcaag ggcttcagcc tccttgctac
     2101 agaggataag gaagccctga agaagcagct cccaggagtc aagagtgaag gaaagagaaa
     2161 aggcgatgag gtggatggag tggatgaagt ggcgaagaag aaatctaaaa aagaaaaaga
     2221 caaggatagt aagcttgaaa aagccctaaa ggctcagaac gacctgatct ggaacatcaa
     2281 ggacgagcta aagaaagtgt gttcaactaa tgacctgaag gagctactca tcttcaacaa
     2341 gcagcaagtg ccttctgggg agtcggcgat cttggaccga gtagctgatg gcatggtgtt
     2401 cggtgccctc cttccctgcg aggaatgctc gggtcagctg gtcttcaaga gcgatgccta
     2461 ttactgcact ggggacgtca ctgcctggac caagtgtatg gtcaagacac agacacccaa
     2521 ccggaaggag tgggtaaccc caaaggaatt ccgagaaatc tcttacctca agaaattgaa
     2581 ggttaaaaag caggaccgta tattcccccc agaaaccagc gcctccgtgg cggccacgcc
     2641 tccgccctcc acagcctcgg ctcctgctgc tgtgaactcc tctgcttcag cagataagcc
     2701 attatccaac atgaagatcc tgactctcgg gaagctgtcc cggaacaagg atgaagtgaa
     2761 ggccatgatt gagaaactcg gggggaagtt gacggggacg gccaacaagg cttccctgtg
     2821 catcagcacc aaaaaggagg tggaaaagat gaataagaag atggaggaag taaaggaagc
     2881 caacatccga gttgtgtctg aggacttcct ccaggacgtc tccgcctcca ccaagagcct
     2941 tcaggagttg ttcttagcgc acatcttgtc cccttggggg gcagaggtga aggcagagcc
     3001 tgttgaagtt gtggccccaa gagggaagtc aggggctgcg ctctccaaaa aaagcaaggg
     3061 ccaggtcaag gaggaaggta tcaacaaatc tgaaaagaga atgaaattaa ctcttaaagg
     3121 aggagcagct gtggatcctg attctggact ggaacactct gcgcatgtcc tggagaaagg
     3181 tgggaaggtc ttcagtgcca cccttggcct ggtggacatc gttaaaggaa ccaactccta
     3241 ctacaagctc cagcttctgg aggacgacaa ggaaaacagg tattggatat tcaggtcctg
     3301 gggccgtgtg ggtacggtga tcggtagcaa caaactggaa cagatgccgt ccaaggagga
     3361 tgccattgag cacttcatga aattatatga agaaaaaacc gggaacgctt ggcactccaa
     3421 aaatttcacg aagtatccca aaaagttcta ccccctggag attgactatg gccaggatga
     3481 agaggcagtg aagaagctga cagtaaatcc tggctaactc gagcaccacc accaccacca
     3541 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     3601 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     3661 aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     3721 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3781 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3841 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3901 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3961 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4021 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4081 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     4141 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4201 aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc atcgagcatc
     4261 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     4321 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     4381 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     4441 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     4501 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     4561 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     4621 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     4681 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     4741 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     4801 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     4861 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     4921 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     4981 ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt
     5041 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
     5101 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5161 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5221 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5281 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     5341 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5401 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5461 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5521 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5581 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5641 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5701 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5761 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5821 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     5881 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     5941 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
     6001 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     6061 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     6121 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     6181 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     6241 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     6301 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     6361 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     6421 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     6481 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     6541 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     6601 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     6661 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
     6721 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
     6781 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
     6841 gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg
     6901 cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca
     6961 ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg
     7021 ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag
     7081 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     7141 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     7201 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     7261 gaggcggttt gcgta
//
