LOCUS       sequence-226627-        8286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of AceS methylase in streptomycetes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8286)
  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 8286)
  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-aceS
FEATURES             Location/Qualifiers
     source          1..8286
                     /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(1685..1701)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1685..1701)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1698..1720)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1709..1725
                     /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(1733..1739,1740..1757,1758..1763))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1778..1799
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6391..6408)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6562..7150)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6642..6661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7321..8112,8113..8181))
                     /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     7944..7963
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8182..8286)
                     /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 taggatcccg
      781 agggagcaag ccatgaccat cggcacactc atcgacgcgg cggtcggtcc gatagcggag
      841 ggcatgagcc ccaccccgca ccgcgacgcc gaggagctgt acgcgcttgc ggccggcatc
      901 gacgtcgcgg acatcgatcg cgacggcgag ccggacgagc agaccaccga cgagttccag
      961 cgcctcaccc gcagccgcgt ctccggagtg ccgatggcct atctgaccgg tgtggccccg
     1021 ttcgccgggc tggagctacg ggtcggcgca ggcgtcttcg tgccccggcg tgacttcagc
     1081 gacgttgttg agcccgtcct gtcccacctc accggcgtca ccgcgcccgt ggtcgtcgac
     1141 ctgtgcgcgg gcgccggggc gcacgccctg gtgatcgccc accgccgccc cgacgccgag
     1201 gtgcacgcgg tggagatcga cgaggaggcg tacggcttcg ccgagctcaa cgccgccgag
     1261 cgcgtggccg ggggcgacac cggcatccac ctgcaccggg gcgatgtcac gcgtcccggc
     1321 gtgctggccg atctggacgg cacggtggac gtggtggtgt cgaacccgcc gttcgtgccc
     1381 gagcaggcgc agcttccgcc cgagttcgcg gtgcaccagc cgcggatcgc cgtcttcgcc
     1441 ggcggcgacg gcctggaggt gatccgcggc gtcatcgaca ccgcggcccg actgctgcgg
     1501 cccggcggcc ggctcgttct ggaacacggc cacctgcacg gcgagacgat gccggcgctg
     1561 ttcgccgcgg acgaccgctt cacggacatc gcgctgcaca ccgaccagtt cggctacccg
     1621 cactacgtgg tcgccacccg cgtctgatca ggacgcgtcg tccatcaagc ttggcgtaat
     1681 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1741 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     1801 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ggaccggcgc
     1861 cacaccggct tgcgccggct cacggtctcg gacggcgcga agccttcgca agtaccttcg
     1921 tgactacccg gtggacgcgt tctccatgct caaggggatg cgcaaggcga tggaactcat
     1981 cggctacccg ccgacgtcca cgagcgtttt cacagtcgag gcgatcgcgg aaggcgtcag
     2041 cgccatcgag tcgttcaagg gcgtcggccg cgccttcgag gagtgcccgc ccggaccacg
     2101 accgttgacc tcgatatgtc agtggtgatc gctgccgtgt gaggtgatca tcactgacga
     2161 atcgaggtcg aggaaccgag cgtccgagga acagaggcgc ttatcggttg gccgcgagat
     2221 tcctgtcgat cctctcgtgc agcgcgattc cgagggaaac ggaaacgttg agagactcgg
     2281 tctggctcat catggggatg gaaaccgagg cggaagacgc ctcctcgaac aggtcggaag
     2341 gcccaccctt ttcgctgccg aacagcaagg ccagccgatc cggattgtcc ccgagttcct
     2401 tcacggaaat gtcgccatcc gccttgagcg tcatcagctg cataccgctg tcccgaatga
     2461 aggcgatggc ctcctcgcga ccggagagaa cgacgggaag ggagaagacg taacctcggc
     2521 tggccctttg gagacgccgg tccgcgatgc tggtgatgtc actgtcgacc aggatgatcc
     2581 ccgacgctcc gagcgcgagc gacgtgcgta ctatcgcgcc gatgttcccg acgatcttca
     2641 ccccgtcgag aacgacgacg tccccacgcc ggctcgcgat atcgccgaac ctggccgggc
     2701 gagggacgcg ggcgatgccg aatgtcttgg ccttccgctc ccccttgaac aactggttga
     2761 cgatcgagga gtcgatgagg cggaccggta tgttctgccg cccgcacaga tccagcaact
     2821 cagatggaaa aggactgctg tcgctgccgt agacctcgat gaactccacc ccggccgcga
     2881 tgctgtgcat gaggggctcg acgtcctcga tcaacgttgt ctttatgttg gatcgcgacg
     2941 gcttggtgac atcgatgatc cgctgcaccg cgggatcgga cggatttgcg atggtgtcca
     3001 actcagtcat ggtcgtccta ccggctgctg tgttcagtga cgcgattcct ggggtgtgac
     3061 accctacgcg acgatggcgg atggctgccc tgaccggcaa tcaccaacgc aaggggaagt
     3121 cgtcgctctc tggcaaagct ccccgctctt ccccgtccgg gacccgcgcg gtcgatcccc
     3181 gcatatgaag tattcgcctt gatcagtccc ggtggacgcg ccagcggccc gccggagcga
     3241 cggactcccc gacctcgatc gtgtcgccct gagcgtccac gtagacgttg cgtgagagca
     3301 ggactgggcc gccgccgacc gcaccgccct caccaccgac cgcgaccgcg ccatggccgc
     3361 cgccgacggc ctggtcgccg ccgccgcccg ccggttcggc gcctgacccg accaaccccc
     3421 gcgggcgccc ccacccacca ggagaccgac catgaccgac ttcgacggac gcctgaccga
     3481 ggggaccgtg aacctggtcc aggaccccaa cggcggtggc tggtccgccc actgcgctga
     3541 gcccggttgc gactgggccg acttcgccgg accgctcggc ttccagggcc tcgtggccat
     3601 cgctcgccga cacacgcact gaccgcacgt caaagccccg ccggatcacc ggcggggctc
     3661 tcttcggccc tccaagtcac accagcccca aggggcgtcg ggagtggcgg agggaacctc
     3721 tggcccgatt ggtgccagga ttcccaccag accaaagagc aacgggccgg acttcgcacc
     3781 tccgacccgt ccgctcccag actcgcgccc cttagccggg cgagacagga acgttgctcg
     3841 tgcccagagt acggagcgat gccgaggcat tgccagatcg gcccgccggg ccccgctgcc
     3901 actgcgggac cgcaattgcc cacacaccgg gcaaacggcc gcgtatctac tgctcagacc
     3961 gctgccggat ggcagcgaag cgggcgatcg cgcgtgtgac gcgagatgcc gcccgaggca
     4021 aaagcgaaca ccttgggaaa gaaacaacag agtttcccgc acccctccga cctgcggttt
     4081 ctccggacgg ggtggatggg gagagcccga gaggcgacag cctctcggaa gtaggaagca
     4141 cgtcgcggag cgacgctgcc cgactgcgga aagccgcccg gtacagccgc cgccggacgc
     4201 tgtggcggat cagcggggac gccgcgtgca agggctgcgg ccgcgccctg atggaccctg
     4261 cctccggcgt gatcgtcgcc cagacggcgg ccggaacgtc cgtggtcctg ggcctgatgc
     4321 ggtgcgggcg gatctggctc tgcccggtct gcgccgccac gatccggcac aagcgggccg
     4381 aggagatcac cgccgccgtg gtcgagtgga tcaagcgcgg ggggaccgcc tacctggtca
     4441 ccttcacggc ccgccatggg cacacggacc ggctcgcgga cctcatggac gccctccagg
     4501 gcacccggaa gacgccggac agcccccggc ggccgggcgc ctaccagcga ctgatcacgg
     4561 gcggcacgtg ggccggacgc cgggccaagg acgggcaccg ggccgccgac cgcgagggca
     4621 tccgagaccg gatcgggtac gtcggcatga tccgcgcgac cgaagtcacc gtggggcaga
     4681 tcaacggctg gcacccgcac atccacgcga tcgtcctggt cggcggccgg accgaggggg
     4741 agcggtccgc gaagcagatc gtcgccacct tcgagccgac cggcgccgcg ctcgacgagt
     4801 ggcaggggca ctggcggtcc gtgtggaccg ccgccctgcg caaggtcaac cccgccttca
     4861 cgcccgacga ccggcacggc gtcgacttca agcggctgga gaccgagcgc gacgccaacg
     4921 acctcgccga gtacatcgcc aagacccagg acgggaaggc gcccgccctc gaactcgccc
     4981 gcgccgacct caagacggcg accggcggga acgtcgcccc gttcgaactc ctcggacgga
     5041 tcggggacct gaccggcggc atgaccgagg acgacgccgc cggggtcggc tcgctggagt
     5101 ggaacctctc gcgctggcac gagtacgagc gggcaacccg gggacgccgg gccatcgaat
     5161 ggacccgcta cctgcggcag atgctcgggc tcgacggcgg cgacaccgag gccgacgacc
     5221 tcgatctgct cctggcggcc gacgccgacg gcggggagct gcgggccggg gtcgccgtga
     5281 ccgaggacgg atggcacgcg gtcacccgcc gcgccctcga cctcgaggcg acccgggccg
     5341 ccgaaggcaa ggacggcaac gaggattcgg cggccgtggg cgaacgggtg cgggaggtcc
     5401 tggcgctggc cgacgcggcc gacacagtgg tggtgctcac ggcgggggag gtggccgagg
     5461 cgtacgccga catgctcgcc gccctcgccc agcgccgcga ggaagcaact gcacgccgac
     5521 ggcgagagca ggacgacgac caggacgacg acgccgacga ccgccaggag cgggccgccc
     5581 ggcacatcgc ccggctcgca agtgggccca cttcgcacta actcgctccc ccccgccgta
     5641 cgtcatcccg gtgacgtacg gcgggggtcg gtgacgtacg cggcgacggc ggccggggtc
     5701 gaagccgcgg gagtaatcct gggattactc gcccggggtc ggccccgccg gcacttcgtg
     5761 caggcggtac ctcgcgcccg actcgcctcg ctacgagacg tgccgcgtac ggtcgtcggc
     5821 catgagcacc accaccccca gggacgccga cggcgcgaag ctctgcgcct ggtgcggctc
     5881 ggagatcaag caatccggcg tcggccggag ccgggactac tgccgccgct cctgccgcca
     5941 gcgggcgtac gaggcccggc gccagcgcga ggcgatcgtg tccgccgtgg cgtcggcagt
     6001 cgctcgccga gatacgtcac gtgacgaaat gcagcagcct tccattccgt cacgtgacga
     6061 aactcgggcc gcaggtcaga gcacggttcc gcccgctccg tcgtggccgt agtacgagac
     6121 gtcggtgacc gttgccctga ccgcgccgac ggagtcctcg ccggtgaggc ggagttgttc
     6181 cggacggagc aggaccgtgc cggcgcgcag gccgccgggc ggagcggcga ggggcaccgt
     6241 gcccaaggcg gtccgggccg tgtcgccgga gtcgacgtcg acggtgccgg ggagcaggac
     6301 ggcgtcgccg acgaagccgg cgaccctgca ttaatgaatc ggccaacgcg cggggagagg
     6361 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6421 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6481 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6541 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6601 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6661 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6721 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6781 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6841 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6901 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6961 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     7021 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7081 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7141 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7201 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7261 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7321 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7381 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7441 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7501 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7561 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7621 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7681 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7741 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7801 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7861 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7921 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7981 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8041 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8101 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8161 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8221 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8281 tccgcg
//
