LOCUS       Exported                7771 bp ds-DNA     circular SYN 15-DEC-2021
DEFINITION  Bacterial vector for expression of N-terminally 6xHis-tagged 
            proteins with a thrombin site. For other reading frames, use 
            pET-28b(+) or pET-28c(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-28a(+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7771)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Dec 15, 2021 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7771
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     source          164..2698
                     /organism="Neisseria lactamica ATCC 23970"
                     /strain="ATCC 23970"
                     /mol_type="genomic DNA"
                     /note="color: #ffffff"
                     /db_xref="taxon:546265"
                     /culture_collection="ATCC:23970"
     source          164..199
                     /organism="Neisseria lactamica ATCC 23970"
                     /strain="ATCC 23970"
                     /mol_type="genomic DNA"
                     /note="color: #ffffff"
                     /db_xref="taxon:546265"
                     /culture_collection="ATCC:23970"
     terminator      26..73
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     misc_feature    158..163
                     /label=MCS
                     /note="multiple cloning site"
                     /note="color: #99ccff"
     gene            complement(164..2698)
                     /locus_tag="DR91_RS04465"
                     /old_locus_tag="DR91_513"
                     /label=DR91_RS04465
                     /note="color: #a6acb3; direction: LEFT"
     gene            complement(164..199)
                     /locus_tag="DR91_RS04450"
                     /old_locus_tag="DR91_510"
                     /label=DR91_RS04450
                     /note="color: #a6acb3; direction: LEFT"
     CDS             complement(164..184)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     CDS             complement(200..2698)
                     /codon_start=1
                     /locus_tag="DR91_RS04465"
                     /old_locus_tag="DR91_513"
                     /product="CRISPR-associated helicase/endonuclease Cas3"
                     /label=DR91_RS04465-cas3
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Protein Homology."
                     /note="color: #993366"
                     /inference="COORDINATES:same species:RefSeq"
                     /protein_id="WP_036469581.1"
                     /translation="NFDYIAHARQDSSKNWHSHPLQKHLQKVAQLAKRFAGRYGSLFAE
                     YAGLLHDLGKFQESFQKYIRNASGFEKENAHLEDVESTKLRKIPHSTAGAKYAVERLNP
                     FFGHLLAYLIAGHHAGLADWYDKGSLKRRLQQADDELAASLSGFVESSLPEDFFPLSDD
                     DLMRDFFAFWEDGAKLEELHIWMRFLFSCLVDADFLDTEAFMNGYADADTAQAAGLRPK
                     FPGLDELHRRYEQYMAQLSEKADKNSSLNQERHAILQQCFSAAETDRTLFSLTVPTGGG
                     KTLASLGFALKHALKFGKKRIIYAIPFTSIIEQNANVFRNALGDDVVLEHHSNLEVKED
                     KETAKTRLATENWDAPLIVTTNVQLFESLFAAKTSRCRKIHNIADSVVILDEAQQLPRD
                     FQKPITDMMRVLARDYGVTFVLCTATQPELGKNIDAFGRTILEGLPDVREIVADKIALS
                     EKLRRVRIKMPPPNGETQSWQKIADEIAARPCVLAVVNTRKHAQKLFAALPSNGIKLHL
                     SANMCATHCSEVIALVRRYLALYRAGSLHKPLWLVSTQLIEAGVDLDFPCVYRAMAGLD
                     SIAQAAGRCNREGKLPQLGEVVVFRAEEGAPSGSLKQGQDITEEMLKAGLLDDPLSPLA
                     FAEYFRRFNGKGDVDKHGITTLLTAEASNENPLAIKFRTAAERFHLIDNQGVALIVPFI
                     PLAHWEKDGSPQIVEANELDDFFRRHLDGVEVSEWQDILDKQRFPQPPDNSFGQTDQPL
                     LPEPFESWFGLLESDPLKHKWVYRKLQRYTITVYEHELKKLPEHAVFSRAGLLVLDKGY
                     YKAVLGADFDDAAWLPENSVL"
     CDS             complement(2699..2701)
                     /codon_start=1
                     /product="start codon"
                     /label=ATG
                     /note="color: #993366"
                     /translation="M"
     RBS             2709..2714
                     /note="ribosome binding site"
                     /note="color: #a6acb3"
     protein_bind    2745..2769
                     /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)."
                     /note="color: #31849b"
     promoter        complement(2770..2788)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     promoter        3097..3174
                     /gene="lacI"
                     /label=lacI promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             3175..4257
                     /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)."
                     /note="color: #993366"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     CDS             5066..5257
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /label=rop
                     /note="color: #993366"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     rep_origin      complement(5687..6275)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             6397..7212
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /note="color: #ccffcc"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(7305..7760)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagcaccttc cgcttcttct
      181 tggggcccac agagcctcct aaaaccgaat tttcaggtag ccaagccgca tcgtcaaaat
      241 ccgcgccaag cacggctttg taatagccct tatctaacac gagcaatccc gctcttgaaa
      301 aaacggcatg ttcaggcaac tttttcagtt cgtgttcgta cacagtaatc gtgtagcgtt
      361 gcagcttgcg gtaaacccat ttgtgtttga gcgggtcgct ttccaacaga ccgaaccagc
      421 tttcaaacgg ctcgggcagc agtggttgat cggtttgccc aaaggagttg tctggcggct
      481 gcggaaagcg ttgtttgtcc aaaatatcct gccattctga aacttcaaca ccatctagat
      541 gtcgtctgaa aaaatcgtcc agctcgtttg cttcgacgat ttgcggactg ccgtcttttt
      601 cccaatgagc caacgggata aacggcacaa tgagtgccac gccttggtta tcaatcaggt
      661 ggaaacgttc ggcagctgtg cggaatttaa ttgccagcgg attttcattt gatgcttctg
      721 ccgtcaaaag cgttgtgata ccgtgtttgt ccacatcacc tttgccgttg aatcggcgga
      781 aatattcggc aaatgccaac ggggaaagcg ggtcatcaag cagccctgct ttcagcatct
      841 cttcggtaat gtcctgcccc tgtttcaggc tgccgctggg cgcgccttct tcggcgcgga
      901 atacgactac ttcgcccaac tgcggcagtt taccttcacg gttgcaccgt cccgccgcct
      961 gggcaatgct gtccagccct gccatcgccc gatacacgca agggaaatcc aaatccacgc
     1021 ctgcttcaat caactgcgtg ctgaccagcc acaagggctt gtgcaggctg cctgcgcgat
     1081 acagtgccaa atatcggcga accaacgcaa tcacttcgct gcagtgtgtg gcgcacatat
     1141 tggcagataa atgtagcttg attccgttag aaggcagggc ggcaaagagt ttttgggcgt
     1201 gttttcgcgt attgaccact gccaaaacac acgggcgcgc ggctatttca tcggcaattt
     1261 tctgccagct ttgcgtttcg ccgtttggcg gcggcatttt gatgcggacg cggcgcagtt
     1321 tttccgataa ggcaattttg tctgccacaa tttcgcgcac atctggtagc ccttccaaaa
     1381 tagtgcgacc gaatgcgtcg atatttttgc caagctccgg ttgggttgcc gtgcacagca
     1441 caaaggtaac gccgtaatca cgcgccagca cccgcatcat gtcggtaatc ggtttttgga
     1501 aatcgcgcgg aagctgctgg gcttcatcca aaatcaccac gctgtcggca atattgtgaa
     1561 tcttgcggca acggctggtt ttcgccgcaa acaggctttc aaacagttgc acattggtag
     1621 taacaatcag cggcgcgtcc caattttccg tagcaagacg agttttcgct gtttccttat
     1681 cttctttcac ttccaaattg ctgtggtgtt ctaaaaccac atcatcgcct aatgcattgc
     1741 ggaaaacatt ggcattctgc tcgataatac tggtgaaagg aatagcatag ataatacgtt
     1801 ttttgccaaa tttcagcgcg tgcttcaaag cgaagcccaa gctcgccaaa gttttaccgc
     1861 caccggtcgg cacggttaaa gaaaacaaag tacggtccgt ttctgcggca gaaaaacatt
     1921 gctgcaaaat ggcgtggcgt tcttggttta aagatgaatt tttatctgct ttttctgaaa
     1981 gttgcgccat atattgctca tatcgccggt gtaactcatc caaaccggga aattttgggc
     2041 gcaatccggc agcctgcgca gtatctgcat cggcatagcc gttcataaag gcttcggtat
     2101 ccaaaaaatc ggcatccacc aagcaggaaa agagaaaacg catccaaata tgcaattctt
     2161 ccagctttgc cccgtcttcc caaaacgcaa aaaaatcccg catcaagtca tcatctgata
     2221 acgggaaaaa atcttcgggc aaactacttt ccacaaagcc cgacaaagac gctgccaact
     2281 cgtcatccgc ctgttgcaga cggcgtttca ggctgccttt gtcataccaa tctgccagcc
     2341 cagcatgatg cccggcaatc aaatatgcca gcaaatgccc gaaaaatgga tttagacgtt
     2401 ctaccgcata tttggcaccg gcagtggaat gcggaatttt gcgcaacttg gtagattcga
     2461 catcttccaa atgggcattt tctttttcaa agccggatgc attacggata tatttctgaa
     2521 aagattcctg aaatttcccc aaatcgtgca aaagccccgc atattcggca aacaacgacc
     2581 cataacgccc tgcaaaacgc ttggcgagtt gggcgacttt ttgtagatgt ttttgcaggg
     2641 gatgggaatg ccaatttttt gatgagtctt ggcgagcgtg ggctatatag tcgaaattca
     2701 tggtatatct ccttcttaaa gttaaacaaa attatttcta gaggggaatt gttatccgct
     2761 cacaattccc ctatagtgag tcgtattaat ttcgcgggat cgagatctcg atcctctacg
     2821 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg
     2881 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg
     2941 gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg
     3001 caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa
     3061 tgcaggagtc gcataaggga gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc
     3121 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa
     3181 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
     3241 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
     3301 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
     3361 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     3421 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     3481 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
     3541 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     3601 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     3661 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     3721 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     3781 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     3841 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     3901 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     3961 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
     4021 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     4081 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     4141 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     4201 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     4261 caacgcaatt aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt
     4321 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
     4381 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt
     4441 cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt
     4501 attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt
     4561 cggcgagaag caggccatta tcgccggcat ggcggcccca cgggtgcgca tgatcgtgct
     4621 cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc
     4681 accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac
     4741 aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc
     4801 ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc
     4861 tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg
     4921 catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc
     4981 agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc ccatgaacag
     5041 aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc ttaacatggc
     5101 ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc tggacgcgga
     5161 tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt accgcagctg
     5221 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5281 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5341 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac
     5401 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat atgcggtgtg
     5461 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
     5521 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5581 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5641 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5701 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5761 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5821 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5881 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5941 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6001 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6061 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6121 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6181 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6241 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6301 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg aacaataaaa
     6361 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     6421 tcttgctcta ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     6481 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat
     6541 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     6601 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     6661 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccgggaaaac agcattccag
     6721 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     6781 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     6841 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     6901 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     6961 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     7021 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     7081 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     7141 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     7201 gagtttttct aagaattaat tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7261 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgaaattg taaacgttaa
     7321 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     7381 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     7441 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     7501 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     7561 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     7621 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     7681 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     7741 tgcgccgcta cagggcgcgt cccattcgcc a
//
