LOCUS       sequence-211601-        5292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses high levels of protein in mycobacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5292)
  AUTHORS   Eitson JL, Medeiros JJ, Hoover AR, Srivastava S, Roybal KT, Ainsa 
            JA, Hansen EJ, Gumbo T, van Oers NS
  TITLE     Mycobacterial shuttle vectors designed for high-level protein 
            expression in infected macrophages.
  JOURNAL   Appl Environ Microbiol. 2012 Oct;78(19):6829-37. doi: 
            10.1128/AEM.01674-12. Epub 2012 Jul 20.
  PUBMED    22820329
REFERENCE   2  (bases 1 to 5292)
  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: 
            "10.1128/AEM.01674-12"; journalName: "Appl Environ Microbiol"; date:
            "2012-10"; volume: "78"; issue: "19"; pages: "6829-37"
COMMENT     Sequence Label: pSUM-Kan-MCS2
FEATURES             Location/Qualifiers
     source          1..5292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(15..20,21..38,39..45)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    53..69
                     /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     58..80
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     77..93
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     77..93
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(182..199)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(182..198)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(191..213)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(578..1372)
                     /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(689..708)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1299..1318
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      4356..4900
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4801..4820
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5017..5034
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5271..5292
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
       61 ggataacaat ttcacacagg aaacagctat gaccatgatt acgaattccc ggggatccaa
      121 gcttctgtca tatgtttttc ctccttataa agttaatcag aattcgagct cgcagcttgg
      181 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
      241 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
      301 gcccttccca acagttgcgc agcctgaatg gcgaatggcg ctgatgtccg gcggtgcttt
      361 tgccgttacg caccaccccg tcagtagctg aacaggaggg acagggggtg ggcgaagaac
      421 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg
      481 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc
      541 gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc
      601 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
      661 cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat
      721 agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca
      781 ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca
      841 tccgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca
      901 gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt
      961 tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat
     1021 cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg
     1081 gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg
     1141 cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tggagttcat
     1201 tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc
     1261 ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc
     1321 tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg
     1381 atcctcatcc tgtctcttga tcagatcttg atcccctgcg ccatcagatc cttggcggca
     1441 agaaagccat ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg
     1501 gcaattccgg ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc catgtaagcc
     1561 cactgcaagc tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag atagcccagt
     1621 agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac gtgttccgct
     1681 tcctttagca gcccttgcgc cctgagtgct tgcggcagcg tgaagctgtg cctagaaata
     1741 acgaggacag tcgcacgacg aagttcttct ggatcgcgcc cgtgctggaa gcactcaacc
     1801 tcgaagcgtg tggttgcgga gccatctagc aaccacacga aacatgcgca acgaaccgcg
     1861 caacgaacaa cgcctagaac tggccctaga tgagctgact cgtatcgttg gtaaacctag
     1921 tttgaccagc atgttttaac tacgttcggt gagctgtcaa cggggcctgt aacggcacaa
     1981 cgaaccgtgc aacgagagtg gccacggatg ccaccacagg cactacaacg gagttcgcca
     2041 cgtacatcac cacaaccacc gattctggcg gtgagctccc cgatattcag cggaaatggc
     2101 ttggtatcga ccaagattcg tagaaccccg tctcgtctgg ctggtattca aaacgggcgc
     2161 aacgaaacac gcaacgagac aggcatggcc caaaccagaa aactagcgtc taccaggact
     2221 tttacctgtc cgacccgttg caacggaacc ccccacggaa cccccgcgac acccgctccc
     2281 caattgcgtt agaacagcgg tggattgtcg gcttcgttgt gggccttttg agccgcttcc
     2341 tgttctgccg cacgctcttt cctcgcccga tagccgagtc gcttaacggt gtccagatgc
     2401 agcccgaaat gtttggccgt ttgcggccaa gagtggccct cgtcgtcgtg ataggcgcgg
     2461 atgcgttcgc ggcgtgcagc ctgctcggcg agccactcgc tgcgttcctg cgccacgagc
     2521 cggacgacgt ggcgttcgga tagtccggtg attcgagcgc cttcggcggc ggtcacgcgc
     2581 cgctttttgc ggacagtcgg ctgccggttg tagccgtcgc tgtagccgtc gctgtagccg
     2641 tcgctcatag caatgcctcc atggctgacg cggactttgc gcgccgcgca actgtgctcg
     2701 ccgccgtgcg cgctgctgcg cccttccgcg agatggccga ctggcgcgca ctgagtgtgg
     2761 cctcgtagac cacgatcccg tccgcccaaa tgcgcgactt ggttgtgatc caacgccaaa
     2821 tgctgttggc gatggcgcgg acctcgctgt ccggtagcgg tccgggacac acgtcgttgc
     2881 acgggaattc ggcgtttcgc gcgtggcact cggcatagat cgcgcggccg agtccgtcca
     2941 cgttccgggt cggcaggtag atccgcatga gggcgggacg ataggcccac aacctgacgg
     3001 aatcgaacag tgcgcaattc cgccctagcg gcgtcggagc cgctttgtac gtggtctgct
     3061 gacgccagcg cggcggtggc atgttcgcgc cgagctcggc ctcgatgtgg ctgagtgtgt
     3121 agagatctga gtggagccat tccgtttccc aggcgatgtg gccggggttt ttggtcatga
     3181 ggcctgagta actgcggtcg ccgtcgacgg cgcgccgaag gccttcggcg cacgccgcca
     3241 tgtatgcgag cggcttacgc cgcgcgtatt cggtgcgtgg aacaggggcg ttgagtgccc
     3301 acactgcgtg tgcgtggccg ttggcgcgat tgcccacgat cgcgttgggc agcggatggg
     3361 acccccgggc gctgagcgct cggagcgctg cgtctggatg gtctacgtcc acgaccagca
     3421 ggtttgccag cgctgttggg ttcgcctcga tgtaccggcg gcctagggcc gacgcgcggc
     3481 tttggcggta gatcccctcg agcagatcgt cgcttgccag cggccagtac ggcagccaga
     3541 gctgctcaaa ttcgtcggcg acgtggctca cgcttggtag tagaccacga ttaatcaccg
     3601 gtgtatggtc cgacacgagc tccaagtcag atatttcgct gaggggccac cccacaactg
     3661 cacactcccc cgctctcccg tcgagccctg gtggtggaac accagcgaca gccgagcacc
     3721 cccaaccacc tgtaccaacc aggaggaaca catgcgtcgt ttcgaggacg tttccgggcc
     3781 gctgagagcc gctgtggcgg ccgtacacgc cgccttagac ccgttagacc ccctgccgcc
     3841 tgaatgcgcg ggtacgagcc acacagcgcc cgaacttacg gagctggtgg gctcacctgg
     3901 ctttatggcg tacgaatcgg ctgtgtgcga cctgttgggc gaggtgaggt acgcgctact
     3961 cacgctggca agggcgacac agccgcccca ccgagcccgc acggccgcgc gcggtgtcaa
     4021 caaccgggtg agtcgtgcac accagcaggt gttcgaggct tggctcgaag tgcaggacat
     4081 cgtggcgaac gccgcccgat gagccgcgcc ttacgctggc tgccagccgt tcgcgggctg
     4141 gttggtgcag cgcgtcgagc ggttagaggc cctgcggtgt tccaccaccg caggcctcgc
     4201 cctttttaag gctgaatttg cttgtctccg aatccaactg gcttgtccaa gggtgtatct
     4261 acgcttaatc caaagttcaa acgaggggat tacacatgac caacttcgat aacgttctcg
     4321 gctcgatatt ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta
     4381 atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga
     4441 gctaccaact ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc
     4501 ctttcagttt agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc
     4561 agtggctgct gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt
     4621 accggataag gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga
     4681 gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca
     4741 gcggaatgac accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag
     4801 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc caccactgat ttgagcgtca
     4861 gatttcgtga tgcttgtcag gggggcggag cctatggaaa aacggctttg ccgcggccct
     4921 ctcacttccc tgttaagtat cttcctggca tcttccagga aatctccgcc ccgttcgtaa
     4981 gccatttccg ctcgccgcag tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg
     5041 gaatatatcc tgtatcacat attctgctga cgcaccggtg cagccttttt tctcctgcca
     5101 catgaagcac ttcactgaca ccctcatcag tgccaacata gtaagccagt atatacactc
     5161 cgctagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     5221 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     5281 tagctcactc at
//
