LOCUS       sequence-410983-        7665 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli-Streptomyces shuttle vector for gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7665)
  TITLE     E.coli-Streptomyces shuttle vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7665)
  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            "Unpublished"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: pGM190oriT
FEATURES             Location/Qualifiers
     source          1..7665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(148..519)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            552..661
                     /note="incP origin of transfer"
     CDS             1609..2403
                     /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(1663..1682)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2273..2292
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      2687..3275
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3176..3195
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3429..3446
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3563..3584
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3599..3604,3605..3622,3623..3629)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3637..3653
                     /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     3642..3664
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3661..3677
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3661..3677
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(3855..3889)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     misc_feature    4060..4116
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      complement(4264..4312)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             6027..7469
                     /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"
ORIGIN
        1 ggccgtcgac gacgacgcgg atcggtcttg ccttgctcgt cggtgatgta cttcaccagc
       61 tccgcgaagt cgctcttctt gatggagcgc atggggacgt gcttggcaat cacgcgcacc
      121 ccccggccgt tttagcggct aaaaaagtca tggctctgcc ctcgggcgga ccacgcccat
      181 catgaccttg ccaagctcgt cctgcttctc ttcgatcttc gccagcaggg cgaggatcgt
      241 ggcatcaccg aaccgcgccg tgcgcgggtc gtcggtgagc cagagtttca gcaggccgcc
      301 caggcggccc aggtcgccat tgatgcgggc cagctcgcgg acgtgctcat agtccacgac
      361 gcccgtgatt ttgtagccct ggccgacggc cagcaggtag gccgacaggc tcatgccggc
      421 cgccgccgcc ttttcctcaa tcgctcttcg ttcgtctgga aggcagtaca ccttgatagg
      481 tgggctgccc ttcctggttg gcttggtttc atcagccatc cgcttgccct catctgttac
      541 gccggcggta gccggccagc ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga
      601 ataagggaca gtgaagaagg aacacccgct cgcgggtggg cctacttcac ctatcctgcc
      661 cggctgacgc cgttggatac accaaggaaa gtctacacga accctttggc aaaatcctgt
      721 atatcgtgcg aaaaaggatg gatataccga aaaaatcgct ataatgaccc cgaagcaggg
      781 ttatgcagcg gaaaagatcc gtcgacctgc gcgtccgcct gcctgctttt ggccggcggg
      841 ccgggccgcg gttgtccgcc gggtcggacc ggtcccttgg tcgaccgatg cctcggccac
      901 gacgatgctg ggcccgaagc ggtacgcgac ccgtgccggg ccgcagcgga tcgcggcagc
      961 aggagctccc cgcccgccac ggctggctgg atggtcgcct cacgcgcccg caggggtgac
     1021 ggcgtgggtg cggcggtcag ccccgtgatc gggcgtcaga cggccgctga gggaccgcga
     1081 ccacccctcc cttcccgcct gccgtctttc accgcccggc cggtccgcag acgccgtacg
     1141 gccaggccac ggaggattac gggcagatgg cgcccacccg gcacccgggc tggggcaccc
     1201 tggacagcac gaaggtccgc cccactcacc ggggcgggcc ttcgcgttga gctagcttca
     1261 cgctgccgca agcactcagg gcgcaagggc tgctaaagga agcggaacac gtagaaagcc
     1321 agtccgcaga aacggtgctg accccggatg aatgtcagct actgggctat ctggacaagg
     1381 gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta
     1441 gactgggcgg ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggt
     1501 aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg
     1561 cgcaggggat caagatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     1621 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     1681 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     1741 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactcca agacgaggca
     1801 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     1861 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     1921 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     1981 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     2041 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     2101 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgga tgcccgacgg cgaggatctc
     2161 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     2221 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     2281 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     2341 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     2401 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     2461 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     2521 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaaaa
     2581 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     2641 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2701 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2761 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2821 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2881 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2941 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3001 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3061 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3121 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3181 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3241 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3301 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3361 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     3421 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     3481 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     3541 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     3601 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     3661 caggaaacag ctatgacatg attacgaatt gtacgtaggc gtcagctacg atgttccggg
     3721 gactgctgat ccggtcagca ggtggaagag ggactggatt ccaaagttct caatgctgct
     3781 tgctgttctt gaatgggggg tcgttgacga cgacatggct cgattggcgc gacaagttgc
     3841 tgcgattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat
     3901 ggagttctga ggtcattact ggaccggatc ggggatctgg gctgagggag ccgacggcac
     3961 gcggcggctc acggcgtggc acgcggaacg tccgggcttg cacctcacgt cacgtgagga
     4021 ggcagcgtgg acggcgtcag agaagggagc ggacatatga agcttgcatg cctgcaggtc
     4081 gactctagag gatccccggg taccgagctc gaattcccca gatctaaagt tttgtcgtct
     4141 ttccagacgt tagtaaatga attttctgta tgaggttttg ctaaacaact ttcaacagtt
     4201 tcagcggagt gagaatagaa aggaacaact aaaggaattg cgaataataa ttttttcacg
     4261 ttgaaaatct ccaaaaaaaa aggctccaaa aggagccttt aattgtatcg gtttatcagc
     4321 ttgctttcga ggtgaatttc ttaaacagct tgataccgat agttgcgccg acaatgacaa
     4381 caaccatcgc ccacgcataa ccgatatatt cggtcgctga ggcttgcagg gagtcaaagg
     4441 ccgcttttgc gggatctcgt cgaaggcact agaggcctat atgcggggat cgaccgcgcg
     4501 ggtcccggac ggggaagagc ggggagcttt gccagagagc gacgacttcc ccttgcgttg
     4561 gtgattgccg gtcagggcag ccatccgcca tcgtcgcgta gggtgtcaca ccccaggaat
     4621 cgcgtcactg aacacagcag ccggtaggac gaccatgact gagttggaca ccatcgcaaa
     4681 tccgtccgat cccgcggtgc agcggatcat cgatgtcacc aagccgtcgc gatccaacat
     4741 aaagacaacg ttgatcgagg acgtcgagcc cctcatgcac agcatcgcgg ccggggtgga
     4801 gttcatcgag gtctacggca gcgacagcag tccttttcca tctgagttgc tggatctgtg
     4861 cgggcggcag aacataccgg tccgcctcat cgactcctcg atcgtcaacc agttgttcaa
     4921 gggggagcgg aaggccaaga cattcggcat cgcccgcgtc cctcgcccgg ccaggttcgg
     4981 cgatatcgcg agccggcgtg gggacgtcgt cgttctcgac ggggtgaaga tcgtcgggaa
     5041 catcggcgcg atagtacgca cgtcgctcgc gctcggagcg tcggggatca tcctggtcga
     5101 cagtgacatc accagcatcg cggaccggcg tctccaaagg gccagccgag gttacgtctt
     5161 ctcccttccc gtcgttctct ccggtcgcga ggaggccatc gccttcattc gggacagcgg
     5221 tatgcagctg atgacgctca aggcggatgg cgacatttcc gtgaaggaac tcggggacaa
     5281 tccggatcgg ctggccttgc tgttcggcag cgaaaagggt gggccttccg acctgttcga
     5341 ggaggcgtct tccgcctcgg tttccatccc catgatgagc cagaccgagt ctctcaacgt
     5401 ttccgtttcc ctcggaatcg cgctgcacga gaggatcgac aggaatctcg cggccaaccg
     5461 ataagcgcct ctgttcctcg gacgctcggt tgctcgacct cgattcgtca gtgatgatca
     5521 acaggaccca gaaatggcac gagcccggga ttggcctccc gggctcgccg cccccgaacc
     5581 ggcgcagcat gccgagtgag gacgtttgct ctgtgacgag cgtacgagag aacagcgcgt
     5641 cgacaccacc gcgccccgcc ggaccgaagt gccactgcgg caccccgatc gtccacatcc
     5701 ccgggaagcg ccccaaaatc tactgctcgg acgcctgccg aaagcgcgca aagcgtctca
     5761 ttgcggacca gtcacgcagc gtagcaagca cgactacagc gggtaatccg gacgcgggct
     5821 ttagtagtaa gggaaaatct ccgtccactg caggtcagag cgctgtaact ggaacgggtt
     5881 ctaccccaaa cgagggtgcc gcgacagccg ggcgggagga cccgcgcgac gagcggttcg
     5941 cccggcggga ccggcaccag acggtgtcgc tgaacgaggc gttcaagggc tgcgggaccc
     6001 ggctcactgc cgggacggcc gagctgatgt ggaagccggg cgaggcgacg tggggcaaca
     6061 cgtgccgctg caacaacgtc cacacctgcc cgtggtgcat gagccgcatc ctggccgtac
     6121 ggggctcgaa cgtgcagctc gccgccgacg gtctcgcgga cgccgggtac gggctgcacc
     6181 ttggcacgaa cacgttgcgc cacttcgagc gcatggcgtt cggcaccgtc cgcaagggaa
     6241 tgcggcacgg cctggtcgcg gtcttgcacg acggctggaa gggcgcgtac ggctcgtccg
     6301 gccgccgctg gcgcacgatg cgcgacgact tcgggatcat cggctacgag cgggccttcg
     6361 aggacacctt cggctggggc tccggctggc acctgcactg gcacacgctc tgggtcaccc
     6421 gcgaggtgct cggcccggac gcccaggccg cgttccgcga cgcactcgcc ggagcctggg
     6481 ccgccggggt cgaatccgcg ggcggctaca ccgtcagcga gacgtgcgac cggcccggtt
     6541 gctcctgtga gggcaagggc cacggcaccg acgtgcgccc actcaacggc gcggacgccg
     6601 ctgacggcga tgcgggcaag caggcccgct acctctacaa ggacggcgac aagaccaagg
     6661 gcggcgtcgc caagatcggc ctggagctgg ccggacagaa cttcaaggcc ggtcgcggcg
     6721 acgaccgcat gggaccgctc gacctgggcg acgcggcggc cgccgagctg cagcggcttc
     6781 gtcggcccgg cccgttcgtc gagaagtacc gcgagcgcga gttcggcgtc ttccaggtcc
     6841 gcaagcacta ccggtcgcag aacctgaacc ggctgatcaa ggaactcggc atccagcagg
     6901 acgtccgcac cgaggaggag atcaccgacg acaccgaggg cctggtcgcc atcgcggtca
     6961 tcccggccta catctggtac cgatacatcg cccgcgtcgc cggacgtcgc ctcgacctca
     7021 tcaaggtcgc cgagacgtac ggcctaccgg gcgtccggcg gctcgtcgag tcctggggcc
     7081 tggtgtgggg caaggacgtc ctggacccgc ccgccccgga agcgccggcc gcgccgggcg
     7141 accttgacgc tgaccagatg aggttcgagg tcatgtccga ggaggaagct gctttccgcg
     7201 aggcgcgccg gaaggccaac gaggcccgca cggaggagct ggccgcttcg ctcgaccgag
     7261 tgcgtcagcc gaagaaggag gcgatccgcc cgacgatcag ccttcggaag cgcctgaagc
     7321 cgaagccggt caccgtcgac gtcaagactc ccccgcccgg cgccgcgtcg cccgtgtgcc
     7381 gccgctgcaa gggcaagctc gccccggtac tccagccctg gggccagcac cccggagact
     7441 gcctccgcgt cgacacagcg gtcgcgtagg gccgctgagg gaccgccatg agaccgccac
     7501 cccggacatc cgggtggcgg tctcatggcg gtcccggcgc gaaagccgag ggtctgcctg
     7561 ccgtgaggtg gccggcggac ctgggttact gctggaatcc cgcgcccagt ccgagcgtcc
     7621 gccggcgctt ttcacctggc tccgggtgct cgtcgatgag cacca
//
