LOCUS       sequence-467313-        6081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of a periplasmically expressed His-tagged 
            Serratia Marcescens NucA (Benzonase).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6081)
  TITLE     Homemade Benzonase
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6081)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pET-29b(+)_NSP4ss_SerM_NucA-His
FEATURES             Location/Qualifiers
     source          1..6081
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     929..948
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(968..1045)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1251..1270
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1358..1377
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1358..1376
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1377..1401
                     /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             1416..1438
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2253..2282
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2295..2312
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2365..2383)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2379..2426
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2463..2918
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2550..2569)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2760..2781
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3011..3826)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3011..3826)
                     /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     3734..3753
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3818..3837
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3948..4536
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4437..4456
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4690..4707
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4722..4864
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4790..4809)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4950..4972
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4966..5157)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5966..6081,1..967))
                     /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 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc
       61 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga
      121 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc
      181 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca
      241 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca
      301 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt
      361 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg
      421 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat
      481 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa
      541 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga
      601 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc
      661 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa
      721 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca
      781 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg
      841 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
      901 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg
      961 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
     1021 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc
     1081 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
     1141 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     1201 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     1261 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     1321 gcgtagagga tcgagatcga tctcgatccc gcgaaattaa tacgactcac tataggggaa
     1381 ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata
     1441 tacatatgaa aaaaattacg gcagccgcag ggttgcttct tctggctgct caaccagcaa
     1501 tggcagacac gttagagtct attgacaact gcgcagtggg ttgtccaacg ggaggaagct
     1561 cgaatgtctc gattgtgcgt catgcctata cattaaacaa caactcgact acaaagttcg
     1621 ccaactgggt tgcttaccac atcactaagg atactcctgc cagcgggaaa acgcgtaact
     1681 ggaaaactga cccagcattg aatcccgcag atactcttgc tccagcggac tatacggggg
     1741 cgaatgctgc acttaaagta gaccgcggac atcaggctcc gttggcttcg ttggcaggcg
     1801 taagcgactg ggagagtctt aattacctta gcaatatcac accgcagaaa agcgacttaa
     1861 atcaaggtgc ctgggctcgt cttgaagacc aagaacgtaa actgattgac cgcgctgata
     1921 tttcaagcgt ttatacggtg acagggcctt tgtatgagcg cgatatggga aaattaccag
     1981 ggacacagaa ggcgcacacc atccccagcg cctattggaa agtgattttc attaataact
     2041 ctccagctgt aaaccactat gcggcatttt tatttgacca gaacacgcct aaaggcgccg
     2101 atttttgcca attccgcgtg actgttgatg aaatcgaaaa acgcaccggc ttaatcatct
     2161 gggctggcct tcccgacgac gtacaagcat ccttaaaaag taagccggga gtacttcccg
     2221 aattgatggg ttgtaagaat tctggtaacg gacaccatca tcatcatcat caccatcacc
     2281 attaatagct cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa
     2341 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     2401 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg
     2461 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2521 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2581 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2641 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2701 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2761 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2821 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2881 taacgcgaat tttaacaaaa tattaacgtt tacaatttca ggtggcactt ttcggggaaa
     2941 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3001 gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     3061 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     3121 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     3181 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     3241 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
     3301 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     3361 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     3421 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     3481 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
     3541 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     3601 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     3661 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     3721 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     3781 ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat
     3841 tactgtttat gtaagcagac agttttattg ttcatgacca aaatccctta acgtgagttt
     3901 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3961 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4021 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4081 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4141 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4201 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4261 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4321 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4381 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4441 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4501 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4561 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4621 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4681 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     4741 cttacgcatc tgtgcggtat ttcacaccgc atatatggtg cactctcagt acaatctgct
     4801 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     4861 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     4921 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4981 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     5041 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     5101 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
     5161 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
     5221 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     5281 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     5341 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
     5401 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
     5461 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
     5521 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
     5581 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc
     5641 gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc
     5701 gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg
     5761 aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat
     5821 gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga
     5881 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     5941 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6001 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     6061 gggtggtttt tcttttcacc a
//
