LOCUS       sequence-222232-        9842 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of PegA and AceS in streptomycetes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9842)
  AUTHORS   Sheehan J, Murphy CD, Caffrey P
  TITLE     New insights into polyene macrolide biosynthesis in Couchioplanes 
            caeruleus.
  JOURNAL   Mol Biosyst. 2017 May 2;13(5):866-873. doi: 10.1039/c7mb00112f.
  PUBMED    28383583
REFERENCE   2  (bases 1 to 9842)
  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"; doi: "10"; journalName: 
            "Mol Biosyst"; date: "2017-05-2- 2"; volume: "13"; issue: "5"; 
            pages: "866-873"
COMMENT     Sequence Label: pIAGO-pegA + aceS
FEATURES             Location/Qualifiers
     source          1..9842
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     68..86
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(124..146)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     246..265
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     459..481
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     473..490
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     474..490
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3241..3257)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3241..3257)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3254..3276)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3265..3281
                     /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)."
     promoter        complement(join(3289..3295,3296..3313,3314..3319))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3334..3355
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7947..7964)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8118..8706)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8198..8217)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8877..9668,9669..9737))
                     /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     9500..9519
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9738..9842)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
       61 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
      121 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
      181 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
      241 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
      301 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
      361 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
      421 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
      481 gacggccagt gaattcgagc tcggtaccag cccgacccga gcacgcgccg gcacgcctgg
      541 tcgatgtcgg accggagttc gaggtacgcg gcttgcaggt ccaggaaggg gacgtccatg
      601 cgagtgtccg ttcgagtggc ggcttgcgcc cgatgctagt cgcggttgat cggcgatcgc
      661 aggtgcacgc ggtcgatctt gacggctggc gagaggtgcg gggaggatct gaccgacgcg
      721 gtccacacgt ggcaccgcga tgctgttgtg ggcacaatcg tgccggttgg taggatctgc
      781 taaggaaatt cggcaccgat cggtcgcagg atcggtgcgg gtgtccttgc agggcaccgg
      841 caaacgcccg agcgtaggag acgagatgac atcgcatcgt cccatcctgt tctgttgcac
      901 ccactccacc ggacaggcca cgtccagcct ggttttggcc ggcgagttcg ccgcccgcgg
      961 cgtcaagaac ctgtggtttg cgtccgacga caaccaccgt gcggcggtcg ccgggctgag
     1021 cggggcgagc gaggtcggct tcgtctcgac gggtccggtc aatccccgtg tggcgccgac
     1081 catgtgggac gacgagacgt atcgcagcat cacccagcgg tcccggtgga aaggcaatcg
     1141 agcacggctg cgccagctca tggacccggt gcaccacgcc gaccggttcc gccagctgga
     1201 cgcggcggtg cagcggatcc agccagccct catggtgatc aacaacctct gtatccacgg
     1261 catccaggtg gcgatgaagc ggggtgtgcc gtatgtcatc accggatcgt cgctggccag
     1321 cgacatctgg cagttcgatc tgccggacga ctacccggtg ccgtactccg gcctgccccg
     1381 gcggatgaac ctgcggcagc ggatcggcaa tcggctcttc cgccaccgca acctgctgtc
     1441 catgctggat ccggcactgc tcaggacggt tggcgccatg ctgaaggcca ccgacgagct
     1501 gggcctggac cgccgccagc tgcagcagcg cgcctggatg gaggaggcgg agctggtgct
     1561 ctgcttctcg gtgttcggcc tggactatcc cttcccccac ccgcccaagt tgcagatggt
     1621 cggcgcgatg gtcccgccgc tgccgcagaa cccggtgcct ggcgagaccg accggtggct
     1681 ggatgcacat ccgtcggtgg tcttcatcgc cttcggcact atcacccggc tcaccaaggc
     1741 ggaggtgcgg gcggtggtcg acgtggcccg gcggctgggc gagcggcacc acgtgctgtg
     1801 gaagttgccc cgtgagcagc agcggttcct gcctcccgcc gcggagctgc ccgccaacct
     1861 tcgcgtggag gactggttgc cctctcaata cgacgtgctc gcccactcga acgtccgggt
     1921 gttcttcggc cacggcggca acaacagctt ccacgagggg atctacttcg gaaagccgtc
     1981 actagtgcgg cccctgtggt ttgactgcct ggaccacgcc gttcgggccg tggacagcgg
     2041 cgtagggctc agtgtcgcgc cgggcaccat ggacccggca gagatccaca gcaagctcac
     2101 cgcgcttctg gacgacacgt cgttccgcgc gcgggccgag cacttcgccc agattcagca
     2161 cgaggccggc ggggtacgcc gggccgccga cctcattctg cgctgcgcgg cggtgacgag
     2221 cgccggccag gacgagcagc cggcggtcca gcggtgatct gcagccgggt gccgccgctc
     2281 ggctgacggc gtcgtcggcg gcggccttcc gtccggctga cgtgccttaa gcttcgaggg
     2341 agcaagccat gaccatcggc acactcatcg acgcggcggt cggtccgata gcggagggca
     2401 tgagccccac cccgcaccgc gacgccgagg agctgtacgc gcttgcggcc ggcatcgacg
     2461 tcgcggacat cgatcgcgac ggcgagccgg acgagcagac caccgacgag ttccagcgcc
     2521 tcacccgcag ccgcgtctcc ggagtgccga tggcctatct gaccggtgtg gccccgttcg
     2581 ccgggctgga gctacgggtc ggcgcaggcg tcttcgtgcc ccggcgtgac ttcagcgacg
     2641 ttgttgagcc cgtcctgtcc cacctcaccg gcgtcaccgc gcccgtggtc gtcgacctgt
     2701 gcgcgggcgc cggggcgcac gccctggtga tcgcccaccg ccgccccgac gccgaggtgc
     2761 acgcggtgga gatcgacgag gaggcgtacg gcttcgccga gctcaacgcc gccgagcgcg
     2821 tggccggggg cgacaccggc atccacctgc accggggcga tgtcacgcgt cccggcgtgc
     2881 tggccgatct ggacggcacg gtggacgtgg tggtgtcgaa cccgccgttc gtgcccgagc
     2941 aggcgcagct tccgcccgag ttcgcggtgc accagccgcg gatcgccgtc ttcgccggcg
     3001 gcgacggcct ggaggtgatc cgcggcgtca tcgacaccgc ggcccgactg ctgcggcccg
     3061 gcggccggct cgttctggaa cacggccacc tgcacggcga gacgatgccg gcgctgttcg
     3121 ccgcggacga ccgcttcacg gacatcgcgc tgcacaccga ccagttcggc tacccgcact
     3181 acgtggtcgc cacccgcgtc tgatcaggac gcgtcgtcca tcaagcttgg cgtaatcatg
     3241 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3301 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3361 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagggac cggcgccaca
     3421 ccggcttgcg ccggctcacg gtctcggacg gcgcgaagcc ttcgcaagta ccttcgtgac
     3481 tacccggtgg acgcgttctc catgctcaag gggatgcgca aggcgatgga actcatcggc
     3541 tacccgccga cgtccacgag cgttttcaca gtcgaggcga tcgcggaagg cgtcagcgcc
     3601 atcgagtcgt tcaagggcgt cggccgcgcc ttcgaggagt gcccgcccgg accacgaccg
     3661 ttgacctcga tatgtcagtg gtgatcgctg ccgtgtgagg tgatcatcac tgacgaatcg
     3721 aggtcgagga accgagcgtc cgaggaacag aggcgcttat cggttggccg cgagattcct
     3781 gtcgatcctc tcgtgcagcg cgattccgag ggaaacggaa acgttgagag actcggtctg
     3841 gctcatcatg gggatggaaa ccgaggcgga agacgcctcc tcgaacaggt cggaaggccc
     3901 acccttttcg ctgccgaaca gcaaggccag ccgatccgga ttgtccccga gttccttcac
     3961 ggaaatgtcg ccatccgcct tgagcgtcat cagctgcata ccgctgtccc gaatgaaggc
     4021 gatggcctcc tcgcgaccgg agagaacgac gggaagggag aagacgtaac ctcggctggc
     4081 cctttggaga cgccggtccg cgatgctggt gatgtcactg tcgaccagga tgatccccga
     4141 cgctccgagc gcgagcgacg tgcgtactat cgcgccgatg ttcccgacga tcttcacccc
     4201 gtcgagaacg acgacgtccc cacgccggct cgcgatatcg ccgaacctgg ccgggcgagg
     4261 gacgcgggcg atgccgaatg tcttggcctt ccgctccccc ttgaacaact ggttgacgat
     4321 cgaggagtcg atgaggcgga ccggtatgtt ctgccgcccg cacagatcca gcaactcaga
     4381 tggaaaagga ctgctgtcgc tgccgtagac ctcgatgaac tccaccccgg ccgcgatgct
     4441 gtgcatgagg ggctcgacgt cctcgatcaa cgttgtcttt atgttggatc gcgacggctt
     4501 ggtgacatcg atgatccgct gcaccgcggg atcggacgga tttgcgatgg tgtccaactc
     4561 agtcatggtc gtcctaccgg ctgctgtgtt cagtgacgcg attcctgggg tgtgacaccc
     4621 tacgcgacga tggcggatgg ctgccctgac cggcaatcac caacgcaagg ggaagtcgtc
     4681 gctctctggc aaagctcccc gctcttcccc gtccgggacc cgcgcggtcg atccccgcat
     4741 atgaagtatt cgccttgatc agtcccggtg gacgcgccag cggcccgccg gagcgacgga
     4801 ctccccgacc tcgatcgtgt cgccctgagc gtccacgtag acgttgcgtg agagcaggac
     4861 tgggccgccg ccgaccgcac cgccctcacc accgaccgcg accgcgccat ggccgccgcc
     4921 gacggcctgg tcgccgccgc cgcccgccgg ttcggcgcct gacccgacca accccccccg
     4981 ccgcccccac ccaccaggag accgaccatg accgacttcg acggacgcct gaccgagggg
     5041 accgtgaacc tggtccagga ccccaacggc ggtggctggt ccgcccactg cgctgagccc
     5101 ggttgcgact gggccgactt cgccggaccg ctcggcttcc agggcctcgt ggccatcgct
     5161 cgccgacaca cgcactgacc gcacgtcaaa gccccgccgg atcaccggcg gggctctctt
     5221 cggccctcca agtcacacca gccccaaggg gcgtcgggag tggcggaggg aacctctggc
     5281 ccgattggtg ccaggattcc caccagacca aagagcaacg ggccggactt cgcacctccg
     5341 acccgtccgc tcccagactc gcgcccctta gccgggcgag acaggaacgt tgctcgtgcc
     5401 cagagtacgg agcgatgccg aggcattgcc agatcggccc gccgggcccc gctgccactg
     5461 cgggaccgca attgcccaca caccgggcaa acggccgcgt atctactgct cagaccgctg
     5521 ccggatggca gcgaagcggg cgatcgcgcg tgtgacgcga gatgccgccc gaggcaaaag
     5581 cgaacacctt gggaaagaaa caacagagtt tcccgcaccc ctccgacctg cggtttctcc
     5641 ggacggggtg gatggggaga gcccgagagg cgacagcctc tcggaagtag gaagcacgtc
     5701 gcggagcgac gctgcccgac tgcggaaagc cgcccggtac agccgccgcc ggacgctgtg
     5761 gcggatcagc ggggacgccg cgtgcaaggg ctgcggccgc gccctgatgg accctgcctc
     5821 cggcgtgatc gtcgcccaga cggcggccgg aacgtccgtg gtcctgggcc tgatgcggtg
     5881 cgggcggatc tggctctgcc cggtctgcgc cgccacgatc cggcacaagc gggccgagga
     5941 gatcaccgcc gccgtggtcg agtggatcaa gcgcgggggg accgcctacc tggtcacctt
     6001 cacggcccgc catgggcaca cggaccggct cgcggacctc atggacgccc tccagggcac
     6061 ccggaagacg ccggacagcc cccggcggcc gggcgcctac cagcgactga tcacgggcgg
     6121 cacgtgggcc ggacgccggg ccaaggacgg gcaccgggcc gccgaccgcg agggcatccg
     6181 agaccggatc gggtacgtcg gcatgatccg cgcgaccgaa gtcaccgtgg ggcagatcaa
     6241 cggctggcac ccgcacatcc acgcgatcgt cctggtcggc ggccggaccg agggggagcg
     6301 gtccgcgaag cagatcgtcg ccaccttcga gccgaccggc gccgcgctcg acgagtggca
     6361 ggggcactgg cggtccgtgt ggaccgccgc cctgcgcaag gtcaaccccg ccttcacgcc
     6421 cgacgaccgg cacggcgtcg acttcaagcg gctggagacc gagcgcgacg ccaacgacct
     6481 cgccgagtac atcgccaaga cccaggacgg gaaggcgccc gccctcgaac tcgcccgcgc
     6541 cgacctcaag acggcgaccg gcgggaacgt cgccccgttc gaactcctcg gacggatcgg
     6601 ggacctgacc ggcggcatga ccgaggacga cgccgccggg gtcggctcgc tggagtggaa
     6661 cctctcgcgc tggcacgagt acgagcgggc aacccgggga cgccgggcca tcgaatggac
     6721 ccgctacctg cggcagatgc tcgggctcga cggcggcgac accgaggccg acgacctcga
     6781 tctgctcctg gcggccgacg ccgacggcgg ggagctgcgg gccggggtcg ccgtgaccga
     6841 ggacggatgg cacgcggtca cccgccgcgc cctcgacctc gaggcgaccc gggccgccga
     6901 aggcaaggac ggcaacgagg attcggcggc cgtgggcgaa cgggtgcggg aggtcctggc
     6961 gctggccgac gcggccgaca cagtggtggt gctcacggcg ggggaggtgg ccgaggcgta
     7021 cgccgacatg ctcgccgccc tcgcccagcg ccgcgaggaa gcaactgcac gccgacggcg
     7081 agagcaggac gacgaccagg acgacgacgc cgacgaccgc caggagcggg ccgcccggca
     7141 catcgcccgg ctcgcaagtg ggcccacttc gcactaactc gctccccccc gccgtacgtc
     7201 atcccggtga cgtacggcgg gggtcggtga cgtacgcggc gacggcggcc ggggtcgaag
     7261 ccgcgggagt aatcctggga ttactcgccc ggggtcggcc ccgccggcac ttcgtgcagg
     7321 cggtacctcg cgcccgactc gcctcgctac gagacgtgcc gcgtacggtc gtcggccatg
     7381 agcaccacca cccccaggga cgccgacggc gcgaagctct gcgcctggtg cggctcggag
     7441 atcaagcaat ccggcgtcgg ccggagccgg gactactgcc gccgctcctg ccgccagcgg
     7501 gcgtacgagg cccggcgcca gcgcgaggcg atcgtgtccg ccgtggcgtc ggcagtcgct
     7561 cgccgagata cgtcacgtga cgaaatgcag cagccttcca ttccgtcacg tgacgaaact
     7621 cgggccgcag gtcagagcac ggttccgccc gctccgtcgt ggccgtagta cgagacgtcg
     7681 gtgaccgttg ccctgaccgc gccgacggag tcctcgccgg tgaggcggag ttgttccgga
     7741 cggagcagga ccgtgccggc gcgcaggccg ccgggcggag cggcgagggg caccgtgccc
     7801 aaggcggtcc gggccgtgtc gccggagtcg acgtcgacgg tgccggggag caggacggcg
     7861 tcgccgacga agccggcgac cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     7921 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     7981 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     8041 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     8101 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     8161 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     8221 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     8281 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     8341 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     8401 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     8461 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     8521 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     8581 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     8641 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     8701 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     8761 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     8821 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     8881 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     8941 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     9001 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     9061 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     9121 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     9181 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     9241 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     9301 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     9361 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     9421 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     9481 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     9541 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     9601 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     9661 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     9721 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     9781 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     9841 cg
//
