LOCUS       Exported                5708 bp DNA     circular SYN 16-SEP-2025
DEFINITION  Bacterial vector for inducible expression of N-terminally 
            10xHis-tagged proteins with a Factor Xa site.
ACCESSION   .
VERSION     .
KEYWORDS    pET-16b
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5708)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5708)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5708)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5708)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5708
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(609..5708,1..608)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        127..145
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    146..170
                     /label=lac operator
                     /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             185..207
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             214..216
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             220..246
                     /label=9xHis
                     /note="9xHis affinity tag"
     CDS             262..273
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     misc_feature    274..290
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     terminator      349..396
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        571..599
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        712..816
                     /label=AmpR promoter
     CDS             817..1674
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1848..2436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2622..2761
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2866..3054)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    4332..4353
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4369..5448)
                     /label=lacI
                     /note="lac repressor"
     promoter        5449..5526
                     /label=lacI promoter
ORIGIN
        1 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
       61 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg
      121 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
      181 taattttgtt taactttaag aaggagatat accatgggcc atcatcatca tcatcatcat
      241 catcatcaca gcagcggcca tatcgaaggt cgtcatatgc tcgaggatcc ggctgctaac
      301 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
      361 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
      421 tatcccgcaa gaggcccggc agtaccggca taaccaagcc tatgcctaca gcatccaggg
      481 tgacggtgcc gaggatgacg atgagcgcat tgttagattt catacacggt gcctgactgc
      541 gttagcaatt taactgtgat aaactaccgc attaaagctt atcgatgata agctgtcaaa
      601 catgagaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
      661 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
      721 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
      781 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
      841 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
      901 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
      961 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     1021 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     1081 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     1141 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     1201 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     1261 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     1321 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     1381 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     1441 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     1501 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     1561 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     1621 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     1681 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     1741 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     1801 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     1861 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     1921 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     1981 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2041 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2101 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2161 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2221 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2281 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2341 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     2401 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     2461 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     2521 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     2581 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     2641 cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg
     2701 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     2761 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     2821 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     2881 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     2941 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     3001 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     3061 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     3121 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     3181 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     3241 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat
     3301 ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa
     3361 accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca
     3421 cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag
     3481 ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggccaggacc caacgctgcc
     3541 cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg atggatatgt tctgccaagg
     3601 gttggtttgc gcattcacag ttctccgcaa gaattgattg gctccaattc ttggagtggt
     3661 gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg gctccatgca
     3721 ccgcgacgca acgcggggag gcagacaagg tatagggcgg cgcctacaat ccatgccaac
     3781 ccgttccatg tgctcgccga ggcggcataa atcgccgtga cgatcagcgg tccaatgatc
     3841 gaagttaggc tggtaagagc cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct
     3901 acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga
     3961 atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc
     4021 gcgtcggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     4081 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     4141 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     4201 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     4261 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     4321 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     4381 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4441 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     4501 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     4561 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     4621 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     4681 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     4741 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     4801 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     4861 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     4921 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     4981 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     5041 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     5101 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     5161 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     5221 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     5281 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     5341 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     5401 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     5461 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     5521 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     5581 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     5641 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     5701 gcccgaag
//
