LOCUS       sequence-223373-        8905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of AmphDI glycosyltansferase in streptomycetes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8905)
  AUTHORS   Stephens N, Rawlings B, Caffrey P
  TITLE     Streptomyces nodosus host strains optimized for polyene 
            glycosylation engineering.
  JOURNAL   Biosci Biotechnol Biochem. 2012;76(2):384-7. doi: 
            10.1271/bbb.110673. Epub 2012 Feb 7.
  PUBMED    22313766
REFERENCE   2  (bases 1 to 8905)
  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: "Biosci 
            Biotechnol Biochem."; date: "2012"; volume: "76(2):384-7. doi"; 
            pages: " 10.1271/bbb.110673. Epub 2012 Feb 7"
COMMENT     Sequence Label: pIAGO-amphDI
FEATURES             Location/Qualifiers
     source          1..8905
                     /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(2305..2321)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2305..2321)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2318..2340)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2329..2345
                     /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(2353..2359,2360..2377,2378..2383))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2398..2419
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7010..7027)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7181..7769)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7261..7280)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7940..8731,8732..8800))
                     /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     8563..8582
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8801..8905)
                     /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 taggatctca
      781 tatggcggcg gcgtcggcga atgcgatcat ccggcgactt tcggtgggtg gacgcgaggc
      841 gtgcgggcct gcgcggtccg ccactttccg gagggtgttg tccatgggcg cgcacaggcg
      901 gccgatcctg ttcgtcagtt atgccgaaag cggtcttctc aatccgctgc tcgtactggc
      961 cgaggaattg tcccggcgtg gtgtggagga tctgtggttc gccacggacg agaaggcgcg
     1021 ggaccagatc gagtcggcgt cggcggacag tgagctgcag ttcgcctcgc tcggtgacac
     1081 cgtgtcgcag atgtcggcgg tcacctggga cgacgagacc tatgccgagg tgacgcagcg
     1141 ctcccgcttc aaggcgcacc gcgcggtcat ccggcactcc ttcgcgcccg agacccgggt
     1201 ggagaagtac cgggcgctgg agaaggccgt cgaggagatc cagcccgctc tgatggtgat
     1261 cgagagcatg tgccagttcg ggtacgagct ggcgatcacc aaggggatcc cgttcgtgct
     1321 cggggtgccg ttcctgccga gcaatgtact gacctcgcat gtgccgttcg ccaagtcgta
     1381 cacgccgtcc gggttcccgg tgccgcactc gggcctgccc ggcaagatgt cgctcgccca
     1441 gcgggtggag aacgagctgt tccgggtgcg cacgctcggg atgttcatga cgaaggagat
     1501 ccgggagatc gtcgaggagg acaaccgggt ccgcggcgag ctgggcatct cccccgaggc
     1561 gcggcagatg atggcgcgca tcgaccacgc cgagcaggtg ttgtgctact cggtggcgga
     1621 gctggactac ccgttcccga tgcacgagaa ggtacggctg gtgggcacgc tggtgccgcc
     1681 gctgccgcag gcgccggacg acgagggcct ctcggactgg ctgacggagc agaagtccgt
     1741 ggtcttcatg ggcttcggca ccatcacccg gctcacccgg gagcaggtcg cctcgctggt
     1801 ggaggtggcc cgccgcctcg agggcgaggg ccaccaggtg ctgtggaagc tgccgagcga
     1861 gcagcagcat ctgctgcccc cggccgagga actgcccgcg aacctgcgga tcgagagctg
     1921 ggtgccctca cagctggacg tgctggcgca cccgaacgtc aaggtgttct tcacgcacgc
     1981 cggcggcaac ggctaccacg agggcctgta cttcggaaag cctctggtgg tacggccgtt
     2041 gtgggtggac tgcgacgacc aggccgtgcg cggccaggac ttcggcgtga gcctgaccgt
     2101 cgaccgtccc gagaccgtgg acacggacga cgtcctcgac aagatcaccc gcgtgctgaa
     2161 cgagtcctcc ttcaccgaac gcgccgagta ctacgccggg ttgctgaagg ccgcgggtgg
     2221 tcgtaccgcc gccgcggacc tgctgctcgg cctccccgtc ctggcaaacg actgaccgag
     2281 gaagtcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2341 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2401 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2461 gtcgtgccag ggaccggcgc cacaccggct tgcgccggct cacggtctcg gacggcgcga
     2521 agccttcgca agtaccttcg tgactacccg gtggacgcgt tctccatgct caaggggatg
     2581 cgcaaggcga tggaactcat cggctacccg ccgacgtcca cgagcgtttt cacagtcgag
     2641 gcgatcgcgg aaggcgtcag cgccatcgag tcgttcaagg gcgtcggccg cgccttcgag
     2701 gagtgcccgc ccggaccacg accgttgacc tcgatatgtc agtggtgatc gctgccgtgt
     2761 gaggtgatca tcactgacga atcgaggtcg aggaaccgag cgtccgagga acagaggcgc
     2821 ttatcggttg gccgcgagat tcctgtcgat cctctcgtgc agcgcgattc cgagggaaac
     2881 ggaaacgttg agagactcgg tctggctcat catggggatg gaaaccgagg cggaagacgc
     2941 ctcctcgaac aggtcggaag gcccaccctt ttcgctgccg aacagcaagg ccagccgatc
     3001 cggattgtcc ccgagttcct tcacggaaat gtcgccatcc gccttgagcg tcatcagctg
     3061 cataccgctg tcccgaatga aggcgatggc ctcctcgcga ccggagagaa cgacgggaag
     3121 ggagaagacg taacctcggc tggccctttg gagacgccgg tccgcgatgc tggtgatgtc
     3181 actgtcgacc aggatgatcc ccgacgctcc gagcgcgagc gacgtgcgta ctatcgcgcc
     3241 gatgttcccg acgatcttca ccccgtcgag aacgacgacg tccccacgcc ggctcgcgat
     3301 atcgccgaac ctggccgggc gagggacgcg ggcgatgccg aatgtcttgg ccttccgctc
     3361 ccccttgaac aactggttga cgatcgagga gtcgatgagg cggaccggta tgttctgccg
     3421 cccgcacaga tccagcaact cagatggaaa aggactgctg tcgctgccgt agacctcgat
     3481 gaactccacc ccggccgcga tgctgtgcat gaggggctcg acgtcctcga tcaacgttgt
     3541 ctttatgttg gatcgcgacg gcttggtgac atcgatgatc cgctgcaccg cgggatcgga
     3601 cggatttgcg atggtgtcca actcagtcat ggtcgtccta ccggctgctg tgttcagtga
     3661 cgcgattcct ggggtgtgac accctacgcg acgatggcgg atggctgccc tgaccggcaa
     3721 tcaccaacgc aaggggaagt cgtcgctctc tggcaaagct ccccgctctt ccccgtccgg
     3781 gacccgcgcg gtcgatcccc gcatatgaag tattcgcctt gatcagtccc ggtggacgcg
     3841 ccagcggccc gccggagcga cggactcccc gacctcgatc gtgtcgccct gagcgtccac
     3901 gtagacgttg cgtgagagca ggactgggcc gccgccgacc gcaccgccct caccaccgac
     3961 cgcgaccgcg ccatggccgc cgccgacggc ctggtcgccg ccgccgcccg ccggttcggc
     4021 gcctgacccg accaaccccc gccccgcccc cacccaccag gagaccgacc atgaccgact
     4081 tcgacggacg cctgaccgag gggaccgtga acctggtcca ggaccccaac ggcggtggct
     4141 ggtccgccca ctgcgctgag cccggttgcg actgggccga cttcgccgga ccgctcggct
     4201 tccagggcct cgtggccatc gctcgccgac acacgcactg accgcacgtc aaagccccgc
     4261 cggatcaccg gcggggctct cttcggccct ccaagtcaca ccagccccaa ggggcgtcgg
     4321 gagtggcgga gggaacctct ggcccgattg gtgccaggat tcccaccaga ccaaagagca
     4381 acgggccgga cttcgcacct ccgacccgtc cgctcccaga ctcgcgcccc ttagccgggc
     4441 gagacaggaa cgttgctcgt gcccagagta cggagcgatg ccgaggcatt gccagatcgg
     4501 cccgccgggc cccgctgcca ctgcgggacc gcaattgccc acacaccggg caaacggccg
     4561 cgtatctact gctcagaccg ctgccggatg gcagcgaagc gggcgatcgc gcgtgtgacg
     4621 cgagatgccg cccgaggcaa aagcgaacac cttgggaaag aaacaacaga gtttcccgca
     4681 cccctccgac ctgcggtttc tccggacggg gtggatgggg agagcccgag aggcgacagc
     4741 ctctcggaag taggaagcac gtcgcggagc gacgctgccc gactgcggaa agccgcccgg
     4801 tacagccgcc gccggacgct gtggcggatc agcggggacg ccgcgtgcaa gggctgcggc
     4861 cgcgccctga tggaccctgc ctccggcgtg atcgtcgccc agacggcggc cggaacgtcc
     4921 gtggtcctgg gcctgatgcg gtgcgggcgg atctggctct gcccggtctg cgccgccacg
     4981 atccggcaca agcgggccga ggagatcacc gccgccgtgg tcgagtggat caagcgcggg
     5041 gggaccgcct acctggtcac cttcacggcc cgccatgggc acacggaccg gctcgcggac
     5101 ctcatggacg ccctccaggg cacccggaag acgccggaca gcccccggcg gccgggcgcc
     5161 taccagcgac tgatcacggg cggcacgtgg gccggacgcc gggccaagga cgggcaccgg
     5221 gccgccgacc gcgagggcat ccgagaccgg atcgggtacg tcggcatgat ccgcgcgacc
     5281 gaagtcaccg tggggcagat caacggctgg cacccgcaca tccacgcgat cgtcctggtc
     5341 ggcggccgga ccgaggggga gcggtccgcg aagcagatcg tcgccacctt cgagccgacc
     5401 ggcgccgcgc tcgacgagtg gcaggggcac tggcggtccg tgtggaccgc cgccctgcgc
     5461 aaggtcaacc ccgccttcac gcccgacgac cggcacggcg tcgacttcaa gcggctggag
     5521 accgagcgcg acgccaacga cctcgccgag tacatcgcca agacccagga cgggaaggcg
     5581 cccgccctcg aactcgcccg cgccgacctc aagacggcga ccggcgggaa cgtcgccccg
     5641 ttcgaactcc tcggacggat cggggacctg accggcggca tgaccgagga cgacgccgcc
     5701 ggggtcggct cgctggagtg gaacctctcg cgctggcacg agtacgagcg ggcaacccgg
     5761 ggacgccggg ccatcgaatg gacccgctac ctgcggcaga tgctcgggct cgacggcggc
     5821 gacaccgagg ccgacgacct cgatctgctc ctggcggccg acgccgacgg cggggagctg
     5881 cgggccgggg tcgccgtgac cgaggacgga tggcacgcgg tcacccgccg cgccctcgac
     5941 ctcgaggcga cccgggccgc cgaaggcaag gacggcaacg aggattcggc ggccgtgggc
     6001 gaacgggtgc gggaggtcct ggcgctggcc gacgcggccg acacagtggt ggtgctcacg
     6061 gcgggggagg tggccgaggc gtacgccgac atgctcgccg ccctcgccca gcgccgcgag
     6121 gaagcaactg cacgccgacg gcgagagcag gacgacgacc aggacgacga cgccgacgac
     6181 cgccaggagc gggccgcccg gcacatcgcc cggctcgcaa gtgggcccac ttcgcactaa
     6241 ctcgctcccc cccgccgtac gtcatcccgg tgacgtacgg cgggggtcgg tgacgtacgc
     6301 ggcgacggcg gccggggtcg aagccgcggg agtaatcctg ggattactcg cccggggtcg
     6361 gccccgccgg cacttcgtgc aggcggtacc tcgcgcccga ctcgcctcgc tacgagacgt
     6421 gccgcgtacg gtcgtcggcc atgagcacca ccacccccag ggacgccgac ggcgcgaagc
     6481 tctgcgcctg gtgcggctcg gagatcaagc aatccggcgt cggccggagc cgggactact
     6541 gccgccgctc ctgccgccag cgggcgtacg aggcccggcg ccagcgcgag gcgatcgtgt
     6601 ccgccgtggc gtcggcagtc gctcgccgag atacgtcacg tgacgaaatg cagcagcctt
     6661 ccattccgtc acgtgacgaa actcgggccg caggtcagag cacggttccg cccgctccgt
     6721 cgtggccgta gtacgagacg tcggtgaccg ttgccctgac cgcgccgacg gagtcctcgc
     6781 cggtgaggcg gagttgttcc ggacggagca ggaccgtgcc ggcgcgcagg ccgccgggcg
     6841 gagcggcgag gggcaccgtg cccaaggcgg tccgggccgt gtcgccggag tcgacgtcga
     6901 cggtgccggg gagcaggacg gcgtcgccga cgaagccggc gaccctgcat taatgaatcg
     6961 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     7021 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     7081 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     7141 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7201 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7261 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7321 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7381 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7441 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7501 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7561 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7621 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7681 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7741 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7801 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7861 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7921 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7981 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     8041 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     8101 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     8161 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8221 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8281 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8341 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8401 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8461 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8521 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8581 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8641 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8701 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8761 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8821 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8881 aaaataaaca aataggggtt ccgcg
//
