LOCUS       sequence-433865-        6117 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Bacterial expression of a periplasmically expressed myc-tagged 
            Serratia Marcescens NucA (Benzonase).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6117)
  TITLE     Homemade Benzonase
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6117)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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            "Unpublished"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-myc
FEATURES             Location/Qualifiers
     source          1..6117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(19..834)
                     /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(19..834)
                     /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     742..761
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     826..845
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      956..1544
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1445..1464
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1698..1715
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1730..1872
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1798..1817)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1958..1980
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1974..2165)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2974..4056)
                     /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"
     primer_bind     4018..4037
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4057..4134)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4340..4359
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4447..4466
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4447..4465
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4466..4490
                     /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             4505..4527
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5342..5371
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5378..5407
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5420..5437
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5490..5508)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5504..5551
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5588..6043
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5675..5694)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5885..5906
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
       61 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
      121 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
      181 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
      241 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
      301 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
      361 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
      421 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
      481 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
      541 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
      601 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
      661 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
      721 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
      781 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc
      841 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac
      901 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
      961 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1021 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1081 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     1141 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1201 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1261 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1321 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1381 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     1441 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     1501 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     1561 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     1621 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     1681 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     1741 attttctcct tacgcatctg tgcggtattt cacaccgcat atatggtgca ctctcagtac
     1801 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     1861 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     1921 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     1981 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     2041 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     2101 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     2161 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     2221 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     2281 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     2341 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     2401 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     2461 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
     2521 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
     2581 cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc
     2641 atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag
     2701 gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg
     2761 ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg
     2821 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
     2881 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
     2941 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3001 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3061 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     3121 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     3181 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     3241 tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     3301 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     3361 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     3421 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     3481 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     3541 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     3601 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     3661 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     3721 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     3781 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     3841 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     3901 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     3961 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     4021 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     4081 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat
     4141 ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
     4201 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     4261 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     4321 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     4381 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg
     4441 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
     4501 taattttgtt taactttaag aaggagatat acatatgaaa aaaattacgg cagccgcagg
     4561 gttgcttctt ctggctgctc aaccagcaat ggcagacacg ttagagtcta ttgacaactg
     4621 cgcagtgggt tgtccaacgg gaggaagctc gaatgtctcg attgtgcgtc atgcctatac
     4681 attaaacaac aactcgacta caaagttcgc caactgggtt gcttaccaca tcactaagga
     4741 tactcctgcc agcgggaaaa cgcgtaactg gaaaactgac ccagcattga atcccgcaga
     4801 tactcttgct ccagcggact atacgggggc gaatgctgca cttaaagtag accgcggaca
     4861 tcaggctccg ttggcttcgt tggcaggcgt aagcgactgg gagagtctta attaccttag
     4921 caatatcaca ccgcagaaaa gcgacttaaa tcaaggtgcc tgggctcgtc ttgaagacca
     4981 agaacgtaaa ctgattgacc gcgctgatat ttcaagcgtt tatacggtga cagggccttt
     5041 gtatgagcgc gatatgggaa aattaccagg gacacagaag gcgcacacca tccccagcgc
     5101 ctattggaaa gtgattttca ttaataactc tccagctgta aaccactatg cggcattttt
     5161 atttgaccag aacacgccta aaggcgccga tttttgccaa ttccgcgtga ctgttgatga
     5221 aatcgaaaaa cgcaccggct taatcatctg ggctggcctt cccgacgacg tacaagcatc
     5281 cttaaaaagt aagccgggag tacttcccga attgatgggt tgtaagaatt ctggtaacgg
     5341 agagcaaaag ctgatctctg aagaagactt agggtctgag cagaagttaa ttagtgaaga
     5401 ggacttataa tagctcgagc accaccacca ccaccactga gatccggctg ctaacaaagc
     5461 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     5521 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc
     5581 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5641 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     5701 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     5761 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     5821 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5881 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     5941 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     6001 aaatttaacg cgaattttaa caaaatatta acgtttacaa tttcaggtgg cacttttcgg
     6061 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtat
//
