LOCUS       sequence-214188-       10315 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Streptomyces/E.coli shuttle expression vector encoding creD and 
            creE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10315)
  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 10315)
  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-creDE
FEATURES             Location/Qualifiers
     source          1..10315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      9..464
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(96..115)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     306..327
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     terminator      complement(688..736)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     primer_bind     complement(4799..4815)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             4835..4846
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(8316..8332)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8402..8419)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8573..9161)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8653..8672)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9332..10123,10124..10192))
                     /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     9955..9974
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10193..10297)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
       61 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
      121 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
      181 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
      241 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
      301 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
      361 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
      421 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttccattc gccattcagg
      481 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagagcttg
      541 gccggatcta aagttttgtc gtctttccag acgttagtaa atgaattttc tgtatgaggt
      601 tttgctaaac aactttcaac agtttcagcg gagtgagaat agaaaggaac aactaaagga
      661 attgcgaata ataatttttt cacgttgaaa atctccaaaa aaaaaggctc caaaaggagc
      721 ctttaattgt atcggtttat cagcttgctt tcgaggtgaa tttcttaaac agcttgatac
      781 cgatagttgc gccgacaatg acaacaacca tcgcccacgc ataaccgata tattcggtcg
      841 ctgaggcttg cagggagtca aaggccgctt ttgcgggatc atcactgacg aatcgaggtc
      901 gaggaaccga gcgtccgagg aacagaggcg cttatcggtt ggccgcgaga ttcctgtcga
      961 tcctctcgtg cagcgcgatt ccgagggaaa cggaaacgtt gagagactcg gtctggctca
     1021 tcatggggat ggaaaccgag gcggaagacg cctcctcgaa caggtcggaa ggcccaccct
     1081 tttcgctgcc gaacagcaag gccagccgat ccggattgtc cccgagttcc ttcacggaaa
     1141 tgtcgccatc cgccttgagc gtcatcagct gcataccgct gtcccgaatg aaggcgatgg
     1201 cctcctcgcg accggagaga acgacgggaa gggagaagac gtaacctcgg ctggcccttt
     1261 ggagacgccg gtccgcgatg ctggtgatgt cactgtcgac caggatgatc cccgacgctc
     1321 cgagcgcgag cgacgtgcgt actatcgcgc cgatgttccc gacgatcttc accccgtcga
     1381 gaacgacgac gtccccacgc cggctcgcga tatcgccgaa cctggccggg cgagggacgc
     1441 gggcgatgcc gaatgtcttg gccttccgct cccccttgaa caactggttg acgatcgagg
     1501 agtcgatgag gcggaccggt atgttctgcc gcccgcacag atccagcaac tcagatggaa
     1561 aaggactgct gtcgctgccg tagacctcga tgaactccac cccggccgcg atgctgtgca
     1621 tgaggggctc gacgtcctcg atcaacgttg tctttatgtt ggatcgcgac ggcttggtga
     1681 catcgatgat ccgctgcacc gcgggatcgg acggatttgc gatggtgtcc aactcagtca
     1741 tggtcgtcct accggctgct gtgttcagtg acgcgattcc tggggtgtga caccctacgc
     1801 gacgatggcg gatggctgcc ctgaccggca atcaccaacg caaggggaag tcgtcgctct
     1861 ctggcaaagc tccccgctct tccccgtccg ggacccgcgc ggtcgatccc cgcatatatg
     1921 aagtattcgc cttgatcagt cccggtggac gcgccagcgg cccgccggag cgacggactc
     1981 cccgacctcg atcgtgtcgc cctgagcgtc cacgtagacg ttgcgtgaga gcaggactgg
     2041 gccgccgccg accgcaccgc cctcaccacc gaccgcgacc gcgccatggc cgccgccgac
     2101 ggcctggtcg ccgccgccgc ccgccggttc ggcgcctgac ccgaccaacc cccgcggggc
     2161 gccggcactt cgtgctggcg ccccgccccc acccaccagg agaccgacca tgaccgactt
     2221 cgacggacgc ctgaccgagg ggaccgtgaa cctggtccag gaccccaacg gcggtggctg
     2281 gtccgcccac tgcgctgagc ccggttgcga ctgggccgac ttcgccggac cgctcggctt
     2341 ccagggcctc gtggccatcg ctcgccgaca cacgcactga ccgcacgtca aagccccgcc
     2401 ggatcaccgg cggggctctc ttcggccctc caagtcacac cagccccaag gggcgtcggg
     2461 agtggcggag ggaacctctg gcccgattgg tgccaggatt cccaccagac caaagagcaa
     2521 cgggccggac ttcgcacctc cgacccgtcc gctcccagac tcgcgcccct tagccgggcg
     2581 agacaggaac gttgctcgtg cccagagtac ggagcgatgc cgaggcattg ccagatcggc
     2641 ccgccgggcc ccgctgccac tgcgggaccg caattgccca cacaccgggc aaacggccgc
     2701 gtatctactg ctcagaccgc tgccggatgg cagcgaagcg ggcgatcgcg cgtgtgacgc
     2761 gagatgccgc ccgaggcaaa agcgaacacc ttgggaaaga aacaacagag tttcccgcac
     2821 ccctccgacc tgcggtttct ccggacgggg tggatgggga gagcccgaga ggcgacagcc
     2881 tctcggaagt aggaagcacg tcgcggagcg acgctgcccg actgcggaaa gccgcccggt
     2941 acagccgccg ccggacgctg tggcggatca gcggggacgc cgcgtgcaag ggctgcggcc
     3001 gcgccctgat ggaccctgcc tccggcgtaa tcgtcgccca gacggcggcc ggaacgtccg
     3061 tggtcctggg cctgatgcgg tgcgggcgga tctggctctg cccggtctgc gccgccacga
     3121 tccggcacaa gcgggccgag gagatcaccg ccgccgtggt cgagtggatc aagcgcgggg
     3181 ggaccgccta cctggtcacc ttcacggccc gccatgggca cacggaccgg ctcgcggacc
     3241 tcatggacgc cctccagggc acccggaaga cgccggacag cccccggcgg ccgggcgcct
     3301 accagcgact gatcacgggc ggcacgtggg ccggacgccg ggccaaggac gggcaccggg
     3361 ccgccgaccg cgagggcatc cgagaccgga tcgggtacgt cggcatgatc cgcgcgaccg
     3421 aagtcaccgt ggggcagatc aacggctggc acccgcacat ccacgcgatc gtcctggtcg
     3481 gcggccggac cgagggggag cggtccgcga agcagatcgt cgccaccttc gagccgaccg
     3541 gcgccgcgct cgacgagtgg caggggcact ggcggtccgt gtggaccgcc gccctgcgca
     3601 aggtcaaccc cgccttcacg cccgacgacc ggcacggcgt cgacttcaag cggctggaga
     3661 ccgagcgcga cgccaacgac ctcgccgagt acatcgccaa gacccaggac gggaaggcgc
     3721 ccgccctcga actcgcccgc gccgacctca agacggcgac cggcgggaac gtcgccccgt
     3781 tcgaactcct cggacggatc ggggacctga ccggcggcat gaccgaggac gacgccgccg
     3841 gggtcggctc gctggagtgg aacctctcgc gctggcacga gtacgagcgg gcaacccggg
     3901 gacgccgggc catcgaatgg acccgctacc tgcggcagat gctcgggctc gacggcggcg
     3961 acaccgaggc cgacgacctc gatctgctcc tggcggccga cgccgacggc ggggagctgc
     4021 gggccggggt cgccgtgacc gaggacggat ggcacgcggt cacccgccgc gccctcgacc
     4081 tcgaggcgac ccgggccgcc gaaggcaagg acggcaacga ggattcggcg gccgtgggcg
     4141 aacgggtgcg ggaggtcctg gcgctggccg acgcggccga cacagtggtg gtgctcacgg
     4201 cgggggaggt ggccgaggcg tacgccgaca tgctcgccgc cctcgcccag cgccgcgagg
     4261 aagcaactgc acgccgacgg cgagagcagg acgacgacca ggacgacgac gccgacgacc
     4321 gccaggagcg ggccgcccgg cacatcgccc ggctcgcaag tgggcccact tcgcactaac
     4381 tcgctccccc ccgccgtacg tcatcccggt gacgtacggc gggggtcggt gacgtacgcg
     4441 gcgacggcgg ccggggtcga agccgcggga gtaatcctgg gattactcgc ccggggtcgg
     4501 ccccgccggc acttcgtgca ggcggtacca gcccgacccg agcacgcgcc ggcacgcctg
     4561 gtcgatgtcg gaccggagtt cgaggtacgc ggcttgcagg tccaggaagg ggacgtccat
     4621 gcgagtgtcc gttcgagtgg cggcttgcgc ccgatgctag tcgcggttga tcggcgatcg
     4681 caggtgcacg cggtcgatct tgacggctgg cgagaggtgc ggggaggatc tgaccgacgc
     4741 ggtccacacg tggcaccgcg atgctgttgt gggcacaatc gtgccggttg gtaggatcga
     4801 tccactagtt ctagaaataa ttttgtttaa cattaaagag gagaaattac atatgaccag
     4861 accccccgca cccccgcccg gcgcccccgg cgccgacgag ctgctcgact gcgggctgct
     4921 gtcccccgtc cgggccggca cccccgtcga agcgctggtc tgcgactccg cctggctcca
     4981 ggcgatgctc gacgcggagg ccgcgctgac ccgcgcccag gccaggaccg gcttcctgcc
     5041 cgccgccgcg gccgaggcga tcaccgcggc ggcgcgggcc gaccggatcg acctgctcgc
     5101 ggtcgcccgg ggcgcccggg agaccgccaa ccccgtggtg gggctggtgg cggcgctcac
     5161 cgccgcggtc cgccgggacg acccggcggc ggccgagtac gtccaccggg gctccaccag
     5221 ccaggacgtg ctggacaccg gggcgatgct cgtcgcccgc cgggcgctgc gcctgatcgg
     5281 cgacgacctg gaccgggcgg ccgacgccct cgcggcgctc gccgccgacc accgcgacac
     5341 ccccatggcc ggccgcacgc tggccctgca cgccgtgccg acgacgttcg ggctgaaggc
     5401 cgctggctgg ctggaactcg tcagcgaggc cgcggggcgg gtggcgcggc tgcgcgacgg
     5461 gctgccgttc tcgctgggcg gcgcggcggg cacgctcgcc gggtacttcg gcgaccgcac
     5521 ggaccggggc gacccggccg tcctcctcga ccggctcctc gacgcctacg cggcggagac
     5581 cgggctcgcc cggccggtgc tgccctggca cgtgctgcgc acccccgtgg ccgatctggc
     5641 cgccgtgctc gcgttcaccg ccggggcgct cgggaagatc gcggtggacg tgcagtcact
     5701 ggcccgcacg gaggtggcgg aagtcgccga gccggccgtg gaggggcgcg gcgcttcctc
     5761 cgcgatgccg cacaaacgca atcccgtcct gtcgacgctg atccgcagcg ctgcgctcca
     5821 agtgcccgcc ctggccacgg ggttgaccca gtgcctggtg tcggaggacg agcggtcggc
     5881 gggcgcctgg cacgcggagt ggcaaccgct gcgcgagtgc ctgcggctga ccgggggcgc
     5941 cgcccgcacc gccgtggagc tcgcggcggg gctggaggtg gacgccgccc ggatgcgggc
     6001 caacctcgac ctcacggacg gccggatcgt ctcggagtcg gtcgcggtgg cgctcacccc
     6061 gctgctcggc aggcaggcgg cgaaggaact gctgacccgg gccgcgttca ccgccggcca
     6121 cgagggccgg acgctgggcg aggtgctcgg cgaactgccc gaactggacg gggtgctgcc
     6181 gaaggagcgg tgggaggcgc tgctcgatcc ggcacgtgcc accggggtcg cgggagcgct
     6241 ggtggacggc gcgctcgccc ggcgccgccc gccggcccgg tgacctagcg gaggagccca
     6301 tatgagcgtc agacggctca ccgtgtgcat cgtcggtgcg ggaccgcgcg gactgtcggt
     6361 gctggaaagg ttctgtgccc acgaacggaa gtccgcctcg cacccggcgg tgacggttca
     6421 cgtcgtggac ccggcccgcc cgggcgcggg ccgggtgtgg cggacgggtc agccgcgcca
     6481 gctgctgatg aacacggtcg cctcgcaggt gaccgtgttc accgacgggt ccgtcgacat
     6541 ggcgggcccg gtggaggccg gccccagcct ccacgagtgg gcgcgcgaac tggccgccct
     6601 cacccccgtg gaggagctgc tcggcggcca cgacgacgcc accctggccg aggcccgggc
     6661 cctgggcgcc gactcgtacc ccacccgggc cttctacggg tgctacctgg aggagatgtt
     6721 ccgccgggtg gtctgcggcg cgcccgccca cctggaggtg cgggtgcacc gctcgaccgc
     6781 cgtctccctg gcggacgaga cgccgggctc cggaggcgcg cagagcctgc tgctcgccga
     6841 cggcacccgg ctggccgggc tcgacgcggt cgtgctcgcg ctcggccacg tccgcgcgga
     6901 ggagccgggc gcccccgacc cgcgcgccgc cgcgctcggc ctggcccact tcccgccggc
     6961 caaccccgcc gacctcgacc tgtccggcat cgcgcccggc acccccgtcc tgctgcgcgg
     7021 gctcggcctg aacttcttcg accacatggc gctgttcacc ctcggccggg gcggcgcctt
     7081 cagccgccgc ccgcacgggc tgcggtacca cccctcgggg ctcgaaccgc ggctctacgc
     7141 cggttcgcgg cgcggggtcc cgtaccacgc gcgcggggag aacgagaagg gcgtggacgg
     7201 gcgccacacc ccgctgctgc tcacccccga gcgcatcgcc gaactgaccg gccggcaccg
     7261 cgagggcccc ggcctgtcgt tcctccggac gctgtggccc ctgatcgccc gcgaggtcga
     7321 atgcgtctac tacggaacgc tgctcgcctc ccgcggccgg gcggccgaac gggacgcctt
     7381 cgtgacggcg tacctggcgg gcggcgacga cacggaccgg ggcggggtgc tggaacgttt
     7441 cggcatcggc ccggccgacc ggtggtgctg ggagcggacc gcgtccccgc acccccggca
     7501 cggcttcacc gggcccgacg ggcaccgccg ctggctgctg gagcacctgg cccaggacgt
     7561 gcgccgggcc agggcgggca atgtgagcga tccgcacaag gccgccctgg acgtgctgcg
     7621 cgacctgcgc aacgagatcc ggctcgtggt ggaccacggc gggctcgacg gcctgtcgca
     7681 ccgggacgac ctcgacggct ggtacaccgg cctgaacgcg ttcctctcga tcggcccgcc
     7741 cgcctcccgc atcgaggaga tggcggccct gatcgaggcg ggcgtgctcg acgtcgtcgg
     7801 ccccggcctg gaggtcgaca tcgacgaggc ggacgccgcg ttcgtcgccc gctccccgct
     7861 ggtccccggc cggccggtgc gcgcccacgt gctgatcgag gcccggctgc cggtgacgga
     7921 cctgcgccgt accgccgatc cgctgttgcg cgacctgctg cgcagcgggc agtgccggtc
     7981 ctaccgcatc cccgccggcc gggcgccgga gggctacgag accggtggcc tcgaagtcac
     8041 ccgtcgcccg taccgcctgg tggacgcgct gggccgggcg cacccgcggc gcttcgcctt
     8101 cggcgtgccc acggaggccg tgcactgggt caccgccgcc ggggcgcggc ccggcgtcaa
     8161 ctcggtcacc ctgggggacg ccgacgcgat agcccacgcc gtcgcctcgc tcacgccggc
     8221 cgccgccccc cggctgcccg cgtacgagga ccccggcgtc cgctgcccct ccgacgaccg
     8281 gctcaccgag gtgacggcat gacctagggg aattggatcc actagttcta gaagctctgc
     8341 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     8401 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     8461 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     8521 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     8581 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     8641 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     8701 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     8761 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     8821 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     8881 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     8941 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     9001 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     9061 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     9121 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     9181 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     9241 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     9301 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     9361 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     9421 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     9481 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     9541 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     9601 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     9661 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     9721 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     9781 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     9841 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     9901 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     9961 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
    10021 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
    10081 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
    10141 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
    10201 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
    10261 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagt
//
