LOCUS       sequence-431003-        7430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of a periplasmically expressed MBP-tagged 
            Serratia Marcescens NucA (Benzonase); also known as c001.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7430)
  TITLE     Homemade Benzonase
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7430)
  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            "Unpublished"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: MBP-Benzonase
FEATURES             Location/Qualifiers
     source          1..7430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     491..510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     744..761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    776..918
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(844..863)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1004..1026
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1020..1211)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        1707..1784
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             1785..2867
                     /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     complement(1804..1823)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(3110..3115,3116..3131,3132..3138)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3146..3162
                     /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)."
     primer_bind     3151..3173
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             join(3232..3309,3310..4407)
                     /codon_start=1
                     /gene="malE"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This gene encodes a signal sequence that can direct 
                     MBP to the bacterial periplasm."
                     /translation="MKIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYNG
                     LAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAE
                     ITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDK
                     ELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLV
                     DLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPS
                     KPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKD
                     PRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     4381..4404
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             4456..4476
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(5225..5242)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5225..5241)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5234..5256)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5420..5437)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5590..5676
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5768..5795
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5814..5905
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5906..5974,5975..6766)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6124..6143)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6808..7321)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7003..7024)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7215..7234
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
       61 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
      121 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
      181 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
      241 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
      301 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
      361 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
      421 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
      481 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      541 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      601 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
      661 cgttatcccc tgattctgtg gataaccgta ttgccgcctt tgagtgagct gataccgctc
      721 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
      781 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatat ggtgcactct
      841 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
      901 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
      961 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     1021 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     1081 tggtcgtgca gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     1141 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     1201 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     1261 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     1321 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     1381 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     1441 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     1501 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     1561 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     1621 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc
     1681 caggacccaa cgctgcccga aattccgaca ccatcgaatg gtgcaaaacc tttcgcggta
     1741 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     1801 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     1861 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     1921 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     1981 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     2041 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     2101 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     2161 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     2221 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     2281 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     2341 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     2401 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     2461 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     2521 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     2581 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     2641 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     2701 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     2761 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     2821 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     2881 aatgtaagtt agctcactca ttaggcacaa ttctcatgtt tgacagctta tcatcgactg
     2941 cacggtgcac caatgcttct ggcgtcaggc agccatcgga agctgtggta tggctgtgca
     3001 ggtcgtaaat cactgcataa ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt
     3061 ttttgcgccg acatcataac ggttctggca aatattctga aatgagctgt tgacaattaa
     3121 tcatcggctc gtataatgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     3181 cagtccgttt aggtgttttc acgagcactt caccaacaag gaccatagca tatgaaaata
     3241 aaaacaggtg cacgcatcct cgcattatcc gcattaacga cgatgatgtt ttccgcctcg
     3301 gctctcgcca aaatcgaaga aggtaaactg gtaatctgga ttaacggcga taaaggctat
     3361 aacggtctcg ctgaagtcgg taagaaattc gagaaagata ccggaattaa agtcaccgtt
     3421 gagcatccgg ataaactgga agagaaattc ccacaggttg cggcaactgg cgatggccct
     3481 gacattatct tctgggcaca cgaccgcttt ggtggctacg ctcaatctgg cctgttggct
     3541 gaaatcaccc cggacaaagc gttccaggac aagctgtatc cgtttacctg ggatgccgta
     3601 cgttacaacg gcaagctgat tgcttacccg atcgctgttg aagcgttatc gctgatttat
     3661 aacaaagatc tgctgccgaa cccgccaaaa acctgggaag agatcccggc gctggataaa
     3721 gaactgaaag cgaaaggtaa gagcgcgctg atgttcaacc tgcaagaacc gtacttcacc
     3781 tggccgctga ttgctgctga cgggggttat gcgttcaagt atgaaaacgg caagtacgac
     3841 attaaagacg tgggcgtgga taacgctggc gcgaaagcgg gtctgacctt cctggttgac
     3901 ctgattaaaa acaaacacat gaatgcagac accgattact ccatcgcaga agctgccttt
     3961 aataaaggcg aaacagcgat gaccatcaac ggcccgtggg catggtccaa catcgacacc
     4021 agcaaagtga attatggtgt aacggtactg ccgaccttca agggtcaacc atccaaaccg
     4081 ttcgttggcg tgctgagcgc aggtattaac gccgccagtc cgaacaaaga gctggcaaaa
     4141 gagttcctcg aaaactatct gctgactgat gaaggtctgg aagcggttaa taaagacaaa
     4201 ccgctgggtg ccgtagcgct gaagtcttac gaggaagagt tggcgaaaga tccacgtatt
     4261 gccgccacta tggaaaacgc ccagaaaggt gaaatcatgc cgaacatccc gcagatgtcc
     4321 gctttctggt atgccgtgcg tactgcggtg atcaacgccg ccagcggtcg tcagactgtc
     4381 gatgaagccc tgaaagacgc gcagactaat tcgagctcga acaacaacaa caataacaat
     4441 aacaacaacc tcggggaaaa cctgtacttc cagggtgata ccctggaaag tattgataac
     4501 tgcgcggtcg gttgtccgac gggtggctct tccaatgtta gcatcgtgcg ccatgcctat
     4561 accctcaaca ataactcaac cactaaattc gcgaattggg tggcgtatca tatcactaag
     4621 gacacgcctg cctcgggtaa gacccgtaac tggaaaaccg acccggcact gaacccagcg
     4681 gatacgcttg caccggctga ctacaccgga gccaatgcag ctctgaaggt ggatcggggc
     4741 caccaggcac cgctggcgtc tcttgcgggc gtaagcgatt gggaatcact gaattatctg
     4801 agcaacatta ccccgcagaa atccgatctg aatcaaggtg cctgggcacg tctggaagac
     4861 caggaacgca aactgattga tcgtgcggat atcagttctg tttacaccgt caccggcccg
     4921 ctgtacgaac gcgatatggg taaactgccg ggcacacaaa aggcgcacac gattccatcg
     4981 gcttattgga aagtgatctt tattaacaat agcccggcag tcaaccatta cgctgcgttc
     5041 ttgtttgacc agaacacccc taaaggcgct gacttttgtc agttccgtgt tactgttgac
     5101 gagatcgaga aacgcactgg attaattatc tgggccggtc tgcccgatga cgtgcaagcg
     5161 tccctcaaga gcaaacccgg ggtattgcca gaattaatgg gttgcaaaaa ctaataagct
     5221 tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     5281 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     5341 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcagcttggc tgttttggcg
     5401 gatgagataa gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa
     5461 aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag
     5521 aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact
     5581 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     5641 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     5701 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
     5761 attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttttgttt
     5821 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5881 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5941 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     6001 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6061 taagatcctt gagagttttc gccccgaaga acgtttccca atgatgagca cttttaaagt
     6121 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     6181 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6241 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6301 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6361 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6421 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6481 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6541 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6601 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6661 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacggaa
     6721 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6781 ttactcatat atactttaga ttgatttacc ccggttgata atcagaaaag ccccaaaaac
     6841 aggaagattg tataagcaaa tatttaaatt gtaaacgtta atattttgtt aaaattcgcg
     6901 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     6961 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     7021 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     7081 ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca
     7141 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     7201 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     7261 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     7321 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     7381 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
//
