LOCUS       sequence-485767-        7260 bp    DNA     circular SYN 11-SEP-2025
DEFINITION  Single vector system expressing hNMT1 (81-496 amino acids) and NEF 
            allowing myristoylation of recombinant protein produce in Ecoli 
            system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7260)
  AUTHORS   Gluck JM, Hoffmann S, Koenig BW, Willbold D
  TITLE     Single vector system for efficient N-myristoylation of recombinant 
            proteins in E. coli.
  JOURNAL   PLoS One. 2010 Apr 9;5(4):e10081. doi: 10.1371/journal.pone.0010081.
  PUBMED    20404920
REFERENCE   2  (bases 1 to 7260)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 11, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2010-04-9"; volume: "5(4):e10081. doi"; pages: " 
            10.1371/journal.pone.0010081"
COMMENT     Sequence Label: pETDuet-1∆HisTag_hNMT_Nef
FEATURES             Location/Qualifiers
     source          1..7260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        1410..1428
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1429..1453
                     /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)."
     CDS             2126..2149
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2150..2167
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2226..2270
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2308..2326)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2322..2369
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2406..2861
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2493..2512)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2703..2724
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2976..3767,3768..3836))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3599..3618
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3837..3929)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4010..4598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4499..4518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4752..4769
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4784..4923
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4849..4868)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5009..5031
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5025..5216)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5788..6870)
                     /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     6832..6851
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6871..6948)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7154..7173
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg aattccttgc cagcagagag gatccaggaa
      121 atacagaagg ccattgagct gttctcagtg ggtcagggac ctgccaaaac catggaggag
      181 gctagcaagc gaagctacca gttctgggat acgcagcccg tccccaagct gggcgaagtg
      241 gtgaacaccc atggccccgt ggagcctgac aaggacaata tccgccagga gccctacacc
      301 ctgccccagg gcttcacctg ggatgctttg gacctgggcg atcgtggtgt gctaaaagaa
      361 ctgtacaccc tcctgaatga gaactatgtg gaagatgatg acaacatgtt ccgatttgat
      421 tattccccgg agtttctttt gtgggctctc cggccacccg gctggctccc ccagtggcac
      481 tgtggggttc gagtggtctc aagtcggaaa ttggttgggt tcattagcgc catcccagca
      541 aacatccata tctatgacac agagaagaag atggtagaga tcaacttcct gtgtgtccac
      601 aagaagctgc gttccaagag ggttgctcca gttctgatcc gagagatcac caggcgggtt
      661 cacctggagg gcatcttcca agcagtttac actgccgggg tggtactacc aaagcctgtt
      721 ggcacctgca ggtattggca tcggtcccta aacccacgga agctgattga agtgaagttc
      781 tcccacctga gcagaaatat gaccatgcag cgcaccatga agctctaccg actgccagag
      841 actcccaaga cagctgggct gcgaccaatg gaaacaaagg acattccagt agtgcaccag
      901 ctcctcacca ggtacttgaa gcaatttcac cttacgcccg tcatgagcca ggaggaggtg
      961 gagcactggt tctaccccca ggagaatatc atcgacactt tcgtggtgga gaacgcaaac
     1021 ggagaggtga cagatttcct gagcttttat acgctgccct ccaccatcat gaaccatcca
     1081 acccacaaga gtctcaaagc tgcttattct ttctacaacg ttcacaccca gacccctctt
     1141 ctagacctca tgagcgacgc ccttgtcctc gccaaaatga aagggtttga tgtgttcaat
     1201 gcactggatc tcatggagaa caaaaccttc ctggagaagc tcaagtttgg cataggggac
     1261 ggcaacctgc agtattacct ttacaattgg aaatgcccca gcatgggggc agagaaggtt
     1321 ggactggtgc tacaataaaa gcttgcggcc gcataatgct taagtcgaac agaaagtaat
     1381 cgtattgtac acggccgcat aatcgaaatt aatacgactc actatagggg aattgtgagc
     1441 ggataacaat tccccatctt agtatattag ttaagtataa gaaggagata tacatatggg
     1501 tggcaagtgg tcaaaacgta gtatgggtgg atggtctgct ataagggaaa gaatgagacg
     1561 agctgagcca cgagctgagc cagcagcaga tggggtggga gcagtatctc gagacctgga
     1621 aaaacatgga gcaatcacaa gtagcaatac agcagctact aatgctgatt gtgcctggct
     1681 agaagcacaa gaggaggaag aggtgggttt tccagtcaga cctcaggtac ctttaagacc
     1741 aatgacttac aaggcagctt tagatattag ccacttttta aaagaaaagg ggggactgga
     1801 agggctaatt tggtcccaaa gaagacaaga gatccttgat ctgtggatct accacacaca
     1861 aggctacttc cctgattggc agaattacac accagggcca gggatcagat atccactgac
     1921 ctttggatgg tgcttcaagc tagtaccagt tgagccagag aaggtagaag aggccaatga
     1981 aggagagaac aacagcttgt tacaccctat gagcctacat gggatggagg acgcggagaa
     2041 agaagtgtta gtgtggaggt ttgacagcaa actagcattt catcacatgg cccgagagct
     2101 gcatccggag tactacaaag actgcctcga agttctgttc caggggcccc atcaccatca
     2161 tcaccactaa caattggata tcggccggcc acgcgatcgc tgacgtcggt accctcgagt
     2221 ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg
     2281 cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     2341 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga
     2401 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2461 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     2521 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2581 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     2641 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2701 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     2761 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     2821 atttaacgcg aattttaaca aaatattaac gtttacaatt tctggcggca cgatggcatg
     2881 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     2941 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3001 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3061 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3121 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3181 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3241 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3301 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3361 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3421 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3481 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3541 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     3601 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     3661 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     3721 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     3781 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     3841 ttcctttttc aatcatgatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3901 atttgaatgt atttagaaaa ataaacaaat aggtcatgac caaaatccct taacgtgagt
     3961 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4021 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4081 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4141 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4201 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4261 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4321 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4381 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4441 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4501 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4561 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4621 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4681 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     4741 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     4801 tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc
     4861 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     4921 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4981 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     5041 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
     5101 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
     5161 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat
     5221 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
     5281 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
     5341 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
     5401 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
     5461 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
     5521 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     5581 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     5641 agccgggtcc tcaacgacag gagcacgatc atgctagtca tgccccgcgc ccaccggaag
     5701 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     5761 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5821 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5881 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     5941 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     6001 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     6061 taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     6121 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     6181 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     6241 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     6301 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     6361 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     6421 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     6481 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     6541 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     6601 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     6661 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     6721 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     6781 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     6841 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     6901 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     6961 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     7021 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     7081 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     7141 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     7201 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat cccgcgaaat
//
