LOCUS       sequence-226848-        7549 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E.coli-Streptomyces shuttle vector, for gene knock out experiments.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7549)
  TITLE     E.coli-Streptomyces shuttle vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7549)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pGMGUS
FEATURES             Location/Qualifiers
     source          1..7549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             579..2021
                     /codon_start=1
                     /gene="rep"
                     /product="replication initiator protein from the 
                     Streptomyces ghanaensis plasmid pSG5"
                     /label=pSG5 Rep
                     /translation="MWKPGEATWGNTCRCNNVHTCPWCMSRILAVRGSNVQLAADGLAD
                     AGYGLHLGTNTLRHFERMAFGTVRKGMRHGLVAVLHDGWKGAYGSSGRRWRTMRDDFGI
                     IGYERAFEDTFGWGSGWHLHWHTLWVTREVLGPDAQAAFRDALAGAWAAGVESAGGYTV
                     SETCDRPGCSCEGKGHGTDVRPLNGADAADGDAGKQARYLYKDGDKTKGGVAKIGLELA
                     GQNFKAGRGDDRMGPLDLGDAAAAELQRLRRPGPFVEKYREREFGVFQVRKHYRSQNLN
                     RLIKELGIQQDVRTEEEITDDTEGLVAIAVIPAYIWYRYIARVAGRRLDLIKVAETYGL
                     PGVRRLVESWGLVWGKDVLDPPAPEAPAAPGDLDADQMRFEVMSEEEAAFREARRKANE
                     ARTEELAASLDRVRQPKKEAIRPTISLRKRLKPKPVTVDVKTPPPGAASPVCRRCKGKL
                     APVLQPWGQHPGDCLRVDTAVA"
     oriT            2424..2533
                     /note="incP origin of transfer"
     CDS             2566..2937
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     CDS             complement(3371..4171)
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     rep_origin      4375..4963
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4864..4883
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5117..5134
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5251..5272
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5287..5292,5293..5310,5311..5317)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5325..5341
                     /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)."
     primer_bind     5330..5352
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5349..5365
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5349..5365
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctagtatcgg acgtctatcg gcgcgtcagg agcgagcagg ggcaatgaac ggccacacca
       61 atgccaggat caacaggacc cagaaatggc acgagcccgg gattggcctc ccgggctcgc
      121 cgcccccgaa ccggcgcagc atgccgagtg aggacgtttg ctctgtgacg agcgtacgag
      181 agaacagcgc gtcgacacca ccgcgccccg ccggaccgaa gtgccactgc ggcaccccga
      241 tcgtccacat ccccgggaag cgccccaaaa tctactgctc ggacgcctgc cgaaagcgcg
      301 caaagcgtct cattgcggac cagtcacgca gcgtagcaag cacgactaca gcgggtaatc
      361 cggacgcggg ctttagtagt aagggaaaat ctccgtccac tgcaggtcag agcgctgtaa
      421 ctggaacggg ttctacccca aacgagggtg ccgcgacagc cgggcgggag gacccgcgcg
      481 acgagcggtt cgcccggcgg gaccggcacc agacggtgtc gctgaacgag gcgttcaagg
      541 gctgcgggac ccggctcact gccgggacgg ccgagctgat gtggaagccg ggcgaggcga
      601 cgtggggcaa cacgtgccgc tgcaacaacg tccacacctg cccgtggtgc atgagccgca
      661 tcctggccgt acggggctcg aacgtgcagc tcgccgccga cggtctcgcg gacgccgggt
      721 acgggctgca ccttggcacg aacacgttgc gccacttcga gcgcatggcg ttcggcaccg
      781 tccgcaaggg aatgcggcac ggcctggtcg cggtcttgca cgacggctgg aagggcgcgt
      841 acggctcgtc cggccgccgc tggcgcacga tgcgcgacga cttcgggatc atcggctacg
      901 agcgggcctt cgaggacacc ttcggctggg gctccggctg gcacctgcac tggcacacgc
      961 tctgggtcac ccgcgaggtg ctcggcccgg acgcccaggc cgcgttccgc gacgcactcg
     1021 ccggagcctg ggccgccggg gtcgaatccg cgggcggcta caccgtcagc gagacgtgcg
     1081 accggcccgg ttgctcctgt gagggcaagg gccacggcac cgacgtgcgc ccactcaacg
     1141 gcgcggacgc cgctgacggc gatgcgggca agcaggcccg ctacctctac aaggacggcg
     1201 acaagaccaa gggcggcgtc gccaagatcg gcctggagct ggccggacag aacttcaagg
     1261 ccggtcgcgg cgacgaccgc atgggaccgc tcgacctggg cgacgcggcg gccgccgagc
     1321 tgcagcggct tcgtcggccc ggcccgttcg tcgagaagta ccgcgagcgc gagttcggcg
     1381 tcttccaggt ccgcaagcac taccggtcgc agaacctgaa ccggctgatc aaggaactcg
     1441 gcatccagca ggacgtccgc accgaggagg agatcaccga cgacaccgag ggcctggtcg
     1501 ccatcgcggt catcccggcc tacatctggt accgatacat cgcccgcgtc gccggacgtc
     1561 gcctcgacct catcaaggtc gccgagacgt acggcctacc gggcgtccgg cggctcgtcg
     1621 agtcctgggg cctggtgtgg ggcaaggacg tcctggaccc gcccgccccg gaagcgccgg
     1681 ccgcgccggg cgaccttgac gctgaccaga tgaggttcga ggtcatgtcc gaggaggaag
     1741 ctgctttccg cgaggcgcgc cggaaggcca acgaggcccg cacggaggag ctggccgctt
     1801 cgctcgaccg agtgcgtcag ccgaagaagg aggcgatccg cccgacgatc agccttcgga
     1861 agcgcctgaa gccgaagccg gtcaccgtcg acgtcaagac tcccccgccc ggcgccgcgt
     1921 cgcccgtgtg ccgccgctgc aagggcaagc tcgccccggt actccagccc tggggccagc
     1981 accccggaga ctgcctccgc gtcgacacag cggtcgcgta gggccgctga gggaccgcca
     2041 tgagaccgcc accccggaca tccgggtggc ggtctcatgg cggtcccggc gcgaaagccg
     2101 agggtctgcc tgccgtgagg tggccggcgg acctgggtta ctgctggaat cccgcgccca
     2161 gtccgagcgt ccgccggcgc ttttcacctg gctccaattg tacatcggat ccatatgacg
     2221 tcgacgcgtc tgcagaagct tcgaattcga gctcccgggt accatggcat gcctgcaggt
     2281 cgacggatct tttccgctgc ataaccctgc ttcggggtca ttatagcgat tttttcggta
     2341 tatccatcct ttttcgcacg atatacagga ttttgccaaa gggttcgtgt agactttcct
     2401 tggtgtatcc aacggcgtca gccgggcagg ataggtgaag taggcccacc cgcgagcggg
     2461 tgttccttct tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg
     2521 cgaggctggc cggctaccgc cggcgtaaca gatgagggca agcggatggc tgatgaaacc
     2581 aagccaacca ggaagggcag cccacctatc aaggtgtact gccttccaga cgaacgaaga
     2641 gcgattgagg aaaaggcggc ggcggccggc atgagcctgt cggcctacct gctggccgtc
     2701 ggccagggct acaaaatcac gggcgtcgtg gactatgagc acgtccgcga gctggcccgc
     2761 atcaatggcg acctgggccg cctgggcggc ctgctgaaac tctggctcac cgacgacccg
     2821 cgcacggcgc ggttcggtga tgccacgatc ctcgccctgc tggcgaagat cgaagagaag
     2881 caggacgagc ttggcaaggt catgatgggc gtggtccgcc cgagggcaga gccatgactt
     2941 ttttagccgc taaaacggcc ggggggtgcg cgtgattgcc aagcacgtcc ccatgcgctc
     3001 catcaagaag agcgacttcg cggagctggt gaagtacatc accgacgagc aaggcaagac
     3061 cgatccccgg ggacctgcag gtcgactcta gctggctggc cagggcccga tacgtcgcgg
     3121 tgagttcagg ctttttcata tctcattgcc cccggacgag cgtctgctcc gccattcgcc
     3181 gtccgccgtg ccaatcggat cagccgtcca aatgcgggat tttcgttagt cggaggccaa
     3241 acggcattga gcgtcagcat atcatcagcg agctgaagaa agacaatccc cgatccgctc
     3301 cacgtgttgc cccagcaatc agcgcgacct tgcccctcca acgtcatctc gttctccgct
     3361 catgagctca gccaatcgac tggcgagcgg catcgcattc ttcgcatccc gcctctggcg
     3421 gatgcaggaa gatcaacgga tctcggccca gttgacccag ggctgtcgcc acaatgtcgc
     3481 gggagcggat caaccgagca aaggcatgac cgactggacc ttccttctga aggctcttct
     3541 ccttgagcca cctgtccgcc aaggcaaagc gctcacagca gtggtcattc tcgagataat
     3601 cgacgcgtac caacttgcca tcctgaagaa tggtgcagtg tctcggcacc ccatagggaa
     3661 cctttgccat caactcggca agatgcagcg tcgtgttggc atcgtgtccc acgccgagga
     3721 gaagtacctg cccatcgagt tcatggacac gggcgaccgg gcttgcaggc gagtgaggtg
     3781 gcaggggcaa tggatcagag atgatctgct ctgcctgtgg ccccgctgcc gcaaaggcaa
     3841 atggatgggc gctgcgcttt acatttggca ggcgccagaa tgtgtcagag acaactccaa
     3901 ggtccggtgt aacgggcgac gtggcaggat cgaacggctc gtcgtccaga cctgaccacg
     3961 agggcatgac gagcgtccct cccggaccca gcgcagcacg cagggcctcg atcagtccaa
     4021 gtggcccatc ttcgaggggc cggacgctac ggaaggagct gtggaccagc agcacaccgc
     4081 cgggggtaac cccaaggttg agaagctgac cgatgagctc ggcttttcgc cattcgtatt
     4141 gcacgacatt gcactccacc gctgatgaca tcagtcgatc atagcacgat caacggcact
     4201 gttgcaaata gtcggtggtg ataaacttat catccccttt tgctgatgga gctgcacatg
     4261 aaccaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4321 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4381 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4441 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcggcag
     4501 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4561 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4621 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4681 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4741 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4801 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4861 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4921 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4981 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5041 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     5101 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     5161 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     5221 ctggaaagcg cgcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     5281 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     5341 atttcacaca ggaaacagct atgacatgat tacgaattgt accgggcccc ccctcgaggt
     5401 ccagcccgac ccgagcacgc gccggcacgc ctggtcgatg tcggaccgga gttcgaggta
     5461 cgcggcttgc aggtccagga aggggacgtc catgcgagtg tccgttcgag tggcggcttg
     5521 cgcccgatgc tagtcgcggt tgatcggcga tcgcaggtgc acgcggtcga tcttgacggc
     5581 tggcgagagg tgcggggagg atctgaccga cgcggtccac acgtggcacc gcgatgctgt
     5641 tgtgggcaca atcgtgccgg ttggtaggat cgacggtatc gataagctag tcgagcaacg
     5701 gaggtacgga catgctgcgg cccgtcgaaa ccccgacccg cgagatcaag aagctggacg
     5761 gcctgtgggc gttcagcctc gaccgggaga actgcggtat cgaccagcgg tggtgggaga
     5821 gcgccctcca ggagtcgcgc gccatcgccg tccccggcag cttcaacgac cagttcgcgg
     5881 acgcggacat ccgcaactac gcggggaacg tctggtatca gcgggaggtg ttcatcccca
     5941 agggctgggc cggtcagcgc atcgtgctgc gcttcgacgc cgtgacccac tacggcaagg
     6001 tctgggtcaa caaccaggaa gtgatggagc accagggcgg ctacacgccc ttcgaggcgg
     6061 acgtgacccc gtacgtcatc gccggcaagt cggtccgcat cacggtctgc gtcaacaacg
     6121 agctcaactg gcagaccatc ccgcccggca tggtgatcac ggacgagaac ggcaagaaga
     6181 agcagagcta cttccacgac ttcttcaact acgccggcat ccaccgctcg gtcatgctgt
     6241 acacgacgcc gaacacctgg gtcgacgaca tcacggtggt cacccacgtg gcccaggact
     6301 gcaaccacgc gtccgtggac tggcaggtcg tcgccaacgg cgacgtgagc gtggagctcc
     6361 gcgacgccga gcagcaggtc gtggcgaccg ggcagggcac ctcggggacc ctccaggtcg
     6421 tgaaccccca cctctggcag ccgggtgagg gctacctgta cgagctgtgc gtgacggcga
     6481 agagccagac cgagtgcgac atctaccccc tgcgcgtcgg catccggtcc gtggcggtca
     6541 agggcgagca gttcctgatc aaccacaagc cgttctactt cacgggcttc ggtcggcacg
     6601 aggacgccga cctccgcggc aagggcttcg acaacgtcct gatggtccac gaccacgccc
     6661 tgatggactg gatcggcgcc aactcgtacc ggacctcgca ctacccgtac gcggaagaga
     6721 tgctggactg ggcggacgag cacgggatcg tcgtcatcga cgaaaccgcc gcggtgggct
     6781 tcaacctgtc cctcggcatc ggcttcgagg ccggcaacaa gccgaaagag ctctactccg
     6841 aagaggccgt caacggcgaa acccagcagg cccacctcca ggcgatcaaa gagctcatcg
     6901 cccgcgacaa gaaccacccg tccgtcgtga tgtggtcgat cgccaacgag ccggacaccc
     6961 ggccgcaggg tgcccgggag tacttcgccc cgctggccga ggccacccgg aagctcgacc
     7021 ccacccggcc catcacctgc gtgaacgtga tgttctgcga cgcccacacg gacaccatct
     7081 ccgacctctt cgacgtcctg tgcctgaacc gctactacgg ctggtacgtg cagtccgggg
     7141 acctggaaac ggccgagaag gtcctggaga aagagctgct ggcgtggcag gagaagctgc
     7201 accagccgat catcatcacc gagtacggcg tggacacgct ggccggtctg cactccatgt
     7261 acaccgacat gtggtccgaa gagtaccagt gcgcgtggct ggacatgtac caccgcgtgt
     7321 tcgaccgcgt cagcgcggtc gtcggcgagc aggtgtggaa cttcgccgac ttcgccacct
     7381 cccagggcat cctccgcgtc gggggcaaca agaagggcat cttcacccgg gaccgcaagc
     7441 ccaagtcggc ggcgttcctg ctccagaagc ggtggaccgg gatgaacttc ggggagaagc
     7501 cgcagcaggg cgggaagcag tgataagcat taccctgtta tccctagtt
//
