LOCUS       sequence-129665-        6539 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli/Streptomyces expression vector containing the T7-promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6539)
  AUTHORS   Muth G
  TITLE     The pSG5-based thermosensitive vector family for genome editing and 
            gene expression in actinomycetes.
  JOURNAL   Appl Microbiol Biotechnol. 2018 Nov;102(21):9067-9080. doi: 
            10.1007/s00253-018-9334-5. Epub 2018 Sep 6.
  PUBMED    30191290
REFERENCE   2  (bases 1 to 6539)
  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: 
            "10.1007/s00253-018-9334-5"; journalName: "Appl Microbiol 
            Biotechnol"; date: "2018-11"; volume: "102"; issue: "21"; pages: 
            "9067-9080"
COMMENT     Sequence Label: pGM202T7
FEATURES             Location/Qualifiers
     source          1..6539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             358..1152
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(412..431)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1022..1041
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      1436..2024
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1925..1944
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2178..2195
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2280..2299
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2280..2298
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2330..2352
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2372..2389
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2393..2425
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(2429..2437,2438..2452)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             2471..2488
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      complement(2680..2728)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             4443..5885
                     /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
                     APVLQPWGRLPGDCLRVDTAVA"
ORIGIN
        1 ctagcttcac gctgccgcaa gcactcaggg cgcaagggct gctaaaggaa gcggaacacg
       61 tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc
      121 tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg
      181 cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg
      241 ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg
      301 atctgatggc gcaggggatc aagatctgat caagagacag gatgaggatc gtttcgcatg
      361 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
      421 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
      481 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactccaa
      541 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
      601 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
      661 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
      721 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
      781 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
      841 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcggat gcccgacggc
      901 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
      961 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     1021 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     1081 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     1141 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     1201 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     1261 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     1321 accccaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     1381 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1441 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1501 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1561 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     1621 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1681 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1741 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1801 ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa
     1861 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     1921 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     1981 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2041 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2101 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     2161 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     2221 aaccgcctct ccccgcgcgt tggccgattc attaatgcag gatctcgatc ccgcgaaatt
     2281 aatacgactc actataggga gaccacaacg gtttccctct agaaataatt ttgtttaact
     2341 ttaagaagga gatatacata tgcggggttc tcatcatcat catcatcatg gtatggctag
     2401 catgactggt ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatccgag
     2461 ctcgagatcc catcatcatc atcatcattg actgcagcca agcttagatc cccgggtacc
     2521 gagctcgaat tccccagatc taaagttttg tcgtctttcc agacgttagt aaatgaattt
     2581 tctgtatgag gttttgctaa acaactttca acagtttcag cggagtgaga atagaaagga
     2641 acaactaaag gaattgcgaa taataatttt ttcacgttga aaatctccaa aaaaaaaggc
     2701 tccaaaagga gcctttaatt gtatcggttt atcagcttgc tttcgaggtg aatttcttaa
     2761 acagcttgat accgatagtt gcgccgacaa tgacaacaac catcgcccac gcataaccga
     2821 tatattcggt cgctgaggct tgcagggagt caaaggccgc ttttgcggga tctcgtcgaa
     2881 ggcactagag gcctatatgc ggggatcgac cgcgcgggtc ccggacgggg aagagcgggg
     2941 agctttgcca gagagcgacg acttcccctt gcgttggtga ttgccggtca gggcagccat
     3001 ccgccatcgt cgcgtagggt gtcacacccc aggaatcgcg tcactgaaca cagcagccgg
     3061 taggacgacc atgactgagt tggacaccat cgcaaatccg tccgatcccg cggtgcagcg
     3121 gatcatcgat gtcaccaagc cgtcgcgatc caacataaag acaacgttga tcgaggacgt
     3181 cgagcccctc atgcacagca tcgcggccgg ggtggagttc atcgaggtct acggcagcga
     3241 cagcagtcct tttccatctg agttgctgga tctgtgcggg cggcagaaca taccggtccg
     3301 cctcatcgac tcctcgatcg tcaaccagtt gttcaagggg gagcggaagg ccaagacatt
     3361 cggcatcgcc cgcgtccctc gcccggccag gttcggcgat atcgcgagcc ggcgtgggga
     3421 cgtcgtcgtt ctcgacgggg tgaagatcgt cgggaacatc ggcgcgatag tacgcacgtc
     3481 gctcgcgctc ggagcgtcgg ggatcatcct ggtcgacagt gacatcacca gcatcgcgga
     3541 ccggcgtctc caaagggcca gccgaggtta cgtcttctcc cttcccgtcg ttctctccgg
     3601 tcgcgaggag gccatcgcct tcattcggga cagcggtatg cagctgatga cgctcaaggc
     3661 ggatggcgac atttccgtga aggaactcgg ggacaatccg gatcggctgg ccttgctgtt
     3721 cggcagcgaa aagggtgggc cttccgacct gttcgaggag gcgtcttccg cctcggtttc
     3781 catccccatg atgagccaga ccgagtctct caacgtttcc gtttccctcg gaatcgcgct
     3841 gcacgagagg atcgacagga atctcgcggc caaccgataa gcgcctctgt tcctcggacg
     3901 ctcggttcct cgacctcgat tcgtcagtga tgatcaacag gacccagaaa tggcacgagc
     3961 ccgggattgg cctcccgggc tcgccgcccc cgaaccggcg cagcatgccg agtgaggacg
     4021 tttgctctgt gacgagcgta cgagagaaca gcgcgtcgac accaccgcgc cccgccggac
     4081 cgaagtgcca ctgcggcacc ccgatcgtcc acatccccgg gaagcgcccc aaaatctact
     4141 gctcggacgc ctgccgaaag cgcgcaaagc gtctcattgc ggaccagtca cgcagcgtag
     4201 caagcacgac tacagcgggt aatccggacg cgggctttag tagtaaggga aaatctccgt
     4261 ccactgcagg tcagagcgct gtaactggaa cgggttctac cccaaacgag ggtgccgcga
     4321 cagccgggcg ggaggacccg cgcgacgagc ggttcgcccg gcgggaccgg caccagacgg
     4381 tgtcgctgaa cgaggcgttc aagggctgcg ggacccggct cactgccggg acggccgagc
     4441 tgatgtggaa gccgggcgag gcgacgtggg gcaacacgtg ccgctgcaac aacgtccaca
     4501 cctgcccgtg gtgcatgagc cgcatcctgg ccgtacgggg ctcgaacgtg cagctcgccg
     4561 ccgacggtct cgcggacgcc gggtacgggc tgcaccttgg cacgaacacg ttgcgccact
     4621 tcgagcgcat ggcgttcggc accgtccgca agggaatgcg gcacggcctg gtcgcggtct
     4681 tgcacgacgg ctggaagggc gcgtacggct cgtccggccg ccgctggcgc acgatgcgcg
     4741 acgacttcgg gatcatcggc tacgagcggg ccttcgagga caccttcggc tggggctccg
     4801 gctggcacct gcactggcac acgctctggg tcacccgcga ggtgctcggc ccggacgccc
     4861 aggccgcgtt ccgcgacgca ctcgccggag cctgggccgc cggggtcgaa tccgcgggcg
     4921 gctacaccgt cagcgagacg tgcgaccggc ccggttgctc ctgtgagggc aagggccacg
     4981 gcaccgacgt gcgcccactc aacggcgcgg acgccgctga cggcgatgcg ggcaagcagg
     5041 cccgctacct ctacaaggac ggcgacaaga ccaagggcgg cgtcgccaag atcggcctgg
     5101 agctggccgg acagaacttc aaggccggtc gcggcgacga ccgcatggga ccgctcgacc
     5161 tgggcgacgc ggcggccgcc gagctgcagc ggcttcgtcg gcccggcccg ttcgtcgaga
     5221 agtaccgcga gcgcgagttc ggcgtcttcc aggtccgcaa gcactaccgg tcgcagaacc
     5281 tgaaccggct gatcaaggaa ctcggcatcc agcaggacgt ccgcaccgag gaggagatca
     5341 ccgacgacac cgagggcctg gtcgccatcg cggtcatccc ggcctacatc tggtaccgat
     5401 acatcgcccg cgtcgccgga cgtcgcctcg acctcatcaa ggtcgccgag acgtacggcc
     5461 taccgggcgt ccggcggctc gtcgagtcct ggggcctggt gtggggcaag gacgtcctgg
     5521 acccgcccgc cccggaagcg ccggccgcgc cgggcgacct tgacgctgac cagatgaggt
     5581 tcgaggtcat gtccgaggag gaagctgctt tccgcgaggc gcgccggaag gccaacgagg
     5641 cccgcacgga ggagctggcc gcttcgctcg accgagtgcg tcagccgaag aaggaggcga
     5701 tccgcccgac gatcagcctt cggaagcgcc tgaagccgaa gccggtcacc gtcgacgtca
     5761 agactccccc gcccggcgcc gcgtcgcccg tgtgccgccg ctgcaagggc aagctcgccc
     5821 cggtactcca gccctggggc cggctccccg gagactgcct ccgcgtcgac acagcggtcg
     5881 cgtagggccg ctgagggacc gccatgagac cgccaccccg gacatccggg tggcggtctc
     5941 atggcggtcc cggcgcgaaa gccgagggtc tgcctgccgt gaggtggccg gcggacctgg
     6001 gttactgctg gaatcccgcg cccagtccga gcgtccgccg gcgcttttca cctggctccg
     6061 ggtgctcgtc gatgagcacc aggccgtcga cgacgacgcg tccgcctgcc tgcttttggc
     6121 cggcgggccg ggccgcggtt gtccgccggg tcggaccggt cccttggtcg accgatgcct
     6181 cggccacgac gatgctgggc ccgaagcggt acgcgaaccc gtgccgggct gcagcggatc
     6241 gcggcagcag gagctccccg cccgccacgg ctggctggat ggtcgcctca cgcgcccgca
     6301 ggggtgacgg cgtgggtgcg gcggtcagcc ccgtgatcgg gcgtcagacg gccgctgagg
     6361 gaccgcgacc acccctccct tcccgcctgc cgtctttcac cgcccggccg gtccgcagac
     6421 gccgtacggc caggccacgg aggattacgg gcagatggcg cccacccggc acccgggctg
     6481 gggcaccctg gacagcacga aggcccgccc cactcaccgg ggcgggcctt cgcgttgag
//
