LOCUS       sequence-214187-        8563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Streptomyces/E.coli shuttle expression vector encoding an N-terminal
            N-His6 tagged CreM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8563)
  AUTHORS   Waldman AJ, Balskus EP
  TITLE     Discovery of a Diazo-Forming Enzyme in Cremeomycin Biosynthesis.
  JOURNAL   J Org Chem. 2018 May 29. doi: 10.1021/acs.joc.8b00367.
  PUBMED    29771512
REFERENCE   2  (bases 1 to 8563)
  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: "J Org Chem.
            2018 May 29. doi: 10.1021/acs.joc.8b00367."
COMMENT     Sequence Label: pUWL201PW-creM-N-His6
FEATURES             Location/Qualifiers
     source          1..8563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      18..473
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(105..124)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     315..336
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     terminator      complement(697..745)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     primer_bind     complement(4808..4824)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             4844..4855
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4865..4882
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6573..6589)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6659..6676)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6830..7418)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6910..6929)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7589..8380,8381..8449))
                     /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     8212..8231
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8450..8554)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
       61 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
      121 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
      181 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
      241 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
      301 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
      361 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
      421 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttccattcg
      481 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
      541 cagagcttgg ccggatctaa agttttgtcg tctttccaga cgttagtaaa tgaattttct
      601 gtatgaggtt ttgctaaaca actttcaaca gtttcagcgg agtgagaata gaaaggaaca
      661 actaaaggaa ttgcgaataa taattttttc acgttgaaaa tctccaaaaa aaaaggctcc
      721 aaaaggagcc tttaattgta tcggtttatc agcttgcttt cgaggtgaat ttcttaaaca
      781 gcttgatacc gatagttgcg ccgacaatga caacaaccat cgcccacgca taaccgatat
      841 attcggtcgc tgaggcttgc agggagtcaa aggccgcttt tgcgggatca tcactgacga
      901 atcgaggtcg aggaaccgag cgtccgagga acagaggcgc ttatcggttg gccgcgagat
      961 tcctgtcgat cctctcgtgc agcgcgattc cgagggaaac ggaaacgttg agagactcgg
     1021 tctggctcat catggggatg gaaaccgagg cggaagacgc ctcctcgaac aggtcggaag
     1081 gcccaccctt ttcgctgccg aacagcaagg ccagccgatc cggattgtcc ccgagttcct
     1141 tcacggaaat gtcgccatcc gccttgagcg tcatcagctg cataccgctg tcccgaatga
     1201 aggcgatggc ctcctcgcga ccggagagaa cgacgggaag ggagaagacg taacctcggc
     1261 tggccctttg gagacgccgg tccgcgatgc tggtgatgtc actgtcgacc aggatgatcc
     1321 ccgacgctcc gagcgcgagc gacgtgcgta ctatcgcgcc gatgttcccg acgatcttca
     1381 ccccgtcgag aacgacgacg tccccacgcc ggctcgcgat atcgccgaac ctggccgggc
     1441 gagggacgcg ggcgatgccg aatgtcttgg ccttccgctc ccccttgaac aactggttga
     1501 cgatcgagga gtcgatgagg cggaccggta tgttctgccg cccgcacaga tccagcaact
     1561 cagatggaaa aggactgctg tcgctgccgt agacctcgat gaactccacc ccggccgcga
     1621 tgctgtgcat gaggggctcg acgtcctcga tcaacgttgt ctttatgttg gatcgcgacg
     1681 gcttggtgac atcgatgatc cgctgcaccg cgggatcgga cggatttgcg atggtgtcca
     1741 actcagtcat ggtcgtccta ccggctgctg tgttcagtga cgcgattcct ggggtgtgac
     1801 accctacgcg acgatggcgg atggctgccc tgaccggcaa tcaccaacgc aaggggaagt
     1861 cgtcgctctc tggcaaagct ccccgctctt ccccgtccgg gacccgcgcg gtcgatcccc
     1921 gcatatatga agtattcgcc ttgatcagtc ccggtggacg cgccagcggc ccgccggagc
     1981 gacggactcc ccgacctcga tcgtgtcgcc ctgagcgtcc acgtagacgt tgcgtgagag
     2041 caggactggg ccgccgccga ccgcaccgcc ctcaccaccg accgcgaccg cgccatggcc
     2101 gccgccgacg gcctggtcgc cgccgccgcc cgccggttcg gcgcctgacc cgaccaaccc
     2161 ccgcggggcg ccggcacttc gtgctggcgc cccgccccca cccaccagga gaccgaccat
     2221 gaccgacttc gacggacgcc tgaccgaggg gaccgtgaac ctggtccagg accccaacgg
     2281 cggtggctgg tccgcccact gcgctgagcc cggttgcgac tgggccgact tcgccggacc
     2341 gctcggcttc cagggcctcg tggccatcgc tcgccgacac acgcactgac cgcacgtcaa
     2401 agccccgccg gatcaccggc ggggctctct tcggccctcc aagtcacacc agccccaagg
     2461 ggcgtcggga gtggcggagg gaacctctgg cccgattggt gccaggattc ccaccagacc
     2521 aaagagcaac gggccggact tcgcacctcc gacccgtccg ctcccagact cgcgcccctt
     2581 agccgggcga gacaggaacg ttgctcgtgc ccagagtacg gagcgatgcc gaggcattgc
     2641 cagatcggcc cgccgggccc cgctgccact gcgggaccgc aattgcccac acaccgggca
     2701 aacggccgcg tatctactgc tcagaccgct gccggatggc agcgaagcgg gcgatcgcgc
     2761 gtgtgacgcg agatgccgcc cgaggcaaaa gcgaacacct tgggaaagaa acaacagagt
     2821 ttcccgcacc cctccgacct gcggtttctc cggacggggt ggatggggag agcccgagag
     2881 gcgacagcct ctcggaagta ggaagcacgt cgcggagcga cgctgcccga ctgcggaaag
     2941 ccgcccggta cagccgccgc cggacgctgt ggcggatcag cggggacgcc gcgtgcaagg
     3001 gctgcggccg cgccctgatg gaccctgcct ccggcgtaat cgtcgcccag acggcggccg
     3061 gaacgtccgt ggtcctgggc ctgatgcggt gcgggcggat ctggctctgc ccggtctgcg
     3121 ccgccacgat ccggcacaag cgggccgagg agatcaccgc cgccgtggtc gagtggatca
     3181 agcgcggggg gaccgcctac ctggtcacct tcacggcccg ccatgggcac acggaccggc
     3241 tcgcggacct catggacgcc ctccagggca cccggaagac gccggacagc ccccggcggc
     3301 cgggcgccta ccagcgactg atcacgggcg gcacgtgggc cggacgccgg gccaaggacg
     3361 ggcaccgggc cgccgaccgc gagggcatcc gagaccggat cgggtacgtc ggcatgatcc
     3421 gcgcgaccga agtcaccgtg gggcagatca acggctggca cccgcacatc cacgcgatcg
     3481 tcctggtcgg cggccggacc gagggggagc ggtccgcgaa gcagatcgtc gccaccttcg
     3541 agccgaccgg cgccgcgctc gacgagtggc aggggcactg gcggtccgtg tggaccgccg
     3601 ccctgcgcaa ggtcaacccc gccttcacgc ccgacgaccg gcacggcgtc gacttcaagc
     3661 ggctggagac cgagcgcgac gccaacgacc tcgccgagta catcgccaag acccaggacg
     3721 ggaaggcgcc cgccctcgaa ctcgcccgcg ccgacctcaa gacggcgacc ggcgggaacg
     3781 tcgccccgtt cgaactcctc ggacggatcg gggacctgac cggcggcatg accgaggacg
     3841 acgccgccgg ggtcggctcg ctggagtgga acctctcgcg ctggcacgag tacgagcggg
     3901 caacccgggg acgccgggcc atcgaatgga cccgctacct gcggcagatg ctcgggctcg
     3961 acggcggcga caccgaggcc gacgacctcg atctgctcct ggcggccgac gccgacggcg
     4021 gggagctgcg ggccggggtc gccgtgaccg aggacggatg gcacgcggtc acccgccgcg
     4081 ccctcgacct cgaggcgacc cgggccgccg aaggcaagga cggcaacgag gattcggcgg
     4141 ccgtgggcga acgggtgcgg gaggtcctgg cgctggccga cgcggccgac acagtggtgg
     4201 tgctcacggc gggggaggtg gccgaggcgt acgccgacat gctcgccgcc ctcgcccagc
     4261 gccgcgagga agcaactgca cgccgacggc gagagcagga cgacgaccag gacgacgacg
     4321 ccgacgaccg ccaggagcgg gccgcccggc acatcgcccg gctcgcaagt gggcccactt
     4381 cgcactaact cgctcccccc cgccgtacgt catcccggtg acgtacggcg ggggtcggtg
     4441 acgtacgcgg cgacggcggc cggggtcgaa gccgcgggag taatcctggg attactcgcc
     4501 cggggtcggc cccgccggca cttcgtgcag gcggtaccag cccgacccga gcacgcgccg
     4561 gcacgcctgg tcgatgtcgg accggagttc gaggtacgcg gcttgcaggt ccaggaaggg
     4621 gacgtccatg cgagtgtccg ttcgagtggc ggcttgcgcc cgatgctagt cgcggttgat
     4681 cggcgatcgc aggtgcacgc ggtcgatctt gacggctggc gagaggtgcg gggaggatct
     4741 gaccgacgcg gtccacacgt ggcaccgcga tgctgttgtg ggcacaatcg tgccggttgg
     4801 taggatcgat ccactagttc tagaaataat tttgtttaac attaaagagg agaaattaca
     4861 tatgcatcat catcatcatc atggcggcgg cgtgcacgca ctgctgcgcc gggtcacacg
     4921 cgggaacggc ttctacatcg gcgacctctt ccacgccgcc gccggccacg acgccaccac
     4981 accggtcacg ctcgaccagc cccttcagta cgcccccgag ctcggcacgc gcttcaccgt
     5041 cgggcagctc gccgagcaga cggacgaact ggccgcccgg ctctgggccg cgggcgtgcg
     5101 gcccaccgaa cgggtcgcgc tgtacaagaa ggacaacttc gacatcgcgc tgcacgcctg
     5161 cgccgccgcg cggatcggcg cggtccccgc cctcctgtcg cccgccctgg aggccccggt
     5221 cgtacggacg ctgctcgacc ggctcgacgg gccgtggctg ctgaccgacg cgacgaccct
     5281 cgccggcccc ctggggacgg tgctcgcacc ggcatccgta cgagcggtcc tgctgacgac
     5341 cggcacaccg gcggagggcg cgcttcccgc agccggcgcg accggacccg ctcgcgaagg
     5401 ggacgcgccg ccgcccgctc ccgtcgtgcg gctggccgac caccggggcg ccccgaagcg
     5461 gccgcccgtc ttcctgcacc cgcgccagcc ctcgctgatc acgcacagct cgggaaccac
     5521 cggcgtcccc aagctggcgg tgcactgccc cgaggcgggc tggcaccggc tggttccgca
     5581 gaaggtcgtg tcctggccga tccgcggcaa ggagaaggcg gcgctgtgca tgacgttcgt
     5641 gcactcgcgc ttctaccagg gcctggccat gttcctcagc cacggcaacc cgatgctgat
     5701 cgccgtcgac cccgacccga gccggatcgg cccgctgttc gcgcgcgaac ggcccggcta
     5761 catcgagacc caccccaaca cctacgtcga ctgggaagcg ctggccgacg ccccgggcga
     5821 accgctcgcc ggcgtacggg tcttcggtgc gaccttcgac gcgatgcacc cccggaccat
     5881 ccagcgcatg ctcggcgcct cccggcgcac ccgcccgctg ttcgtccagt tctacggcca
     5941 gtcggagatc ggcccgatgg ccggccgctg gtacacccgg cgctcggcgg ccaggatgaa
     6001 cggccgctgc gtggggctgc cgctgcccgg cttcatctcg ctgcgggtgg tcgacgacgc
     6061 cgggaagcgg ctgcgcggcg ggcgcaccgg ccacctcgaa gtgcgcagca ggacgcgcat
     6121 cctcacctac ctcggcgagg accagcggta cgcggagcaa ttgcacgacg gatggtggcg
     6181 ggtgggcgac atgggccgcc gcgaccgctg gggcctgctg cacctgctgg accgcgaggt
     6241 cgaccggatc ggcgacctgg agagcagcct ggcggtggag gacctgctga tgagccgcct
     6301 ggaggaactg cgcgaggtgg tcctggtgcc cggggcggac ggcgaaccgg tccccgtcgt
     6361 cgccaccgtc gacgagagcc cgctcgacgc ggcgcgctgg cagcgggcga ccagcgacct
     6421 gccgacgatg gcgccggtgc gccagttccg gttcgaagac ctgccgcgca cgtccacccg
     6481 caagatccag aaaccggaac tggcccggct gatccagggc gtccgggcaa ccgaccaggg
     6541 agtgggggca tgagaattcc tgcagcccgg gggatccact agttctagaa gctctgcatt
     6601 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6661 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6721 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6781 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6841 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6901 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6961 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     7021 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7081 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7141 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7201 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7261 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7321 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7381 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7441 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7501 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7561 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7621 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7681 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7741 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7801 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7861 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7921 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7981 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     8041 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     8101 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     8161 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8221 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8281 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8341 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8401 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8461 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8521 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tcc
//
