LOCUS       sequence-206175-        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"
     CDS             2956..3750
                     /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(3010..3029)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3620..3639
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4115..4137
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4129..4146
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4130..4146
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4235..4251)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4235..4251)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4248..4270)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4259..4275
                     /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(4283..4289,4290..4307,4308..4313))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4328..4349
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4586..4603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4720..5264)
                     /direction=LEFT
                     /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     complement(4800..4819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atcgagccga gaacgttatc gaagttggtc atgtgtaatc ccctcgtttg aactttggat
       61 taagcgtaga tacacccttg gacaagccag ttggattcgg agacaagcaa attcagcctt
      121 aaaaagggcg aggcctgcgg tggtggaaca ccgcagggcc tctaaccgct cgacgcgctg
      181 caccaaccag cccgcgaacg gctggcagcc agcgtaaggc gcggctcatc gggcggcgtt
      241 cgccacgatg tcctgcactt cgagccaagc ctcgaacacc tgctggtgtg cacgactcac
      301 ccggttgttg acaccgcgcg cggccgtgcg ggctcggtgg ggcggctgtg tcgcccttgc
      361 cagcgtgagt agcgcgtacc tcacctcgcc caacaggtcg cacacagccg attcgtacgc
      421 cataaagcca ggtgagccca ccagctccgt aagttcgggc gctgtgtggc tcgtacccgc
      481 gcattcaggc ggcagggggt ctaacgggtc taaggcggcg tgtacggccg ccacagcggc
      541 tctcagcggc ccggaaacgt cctcgaaacg acgcatgtgt tcctcctggt tggtacaggt
      601 ggttgggggt gctcggctgt cgctggtgtt ccaccaccag ggctcgacgg gagagcgggg
      661 gagtgtgcag ttgtggggtg gcccctcagc gaaatatctg acttggagct cgtgtcggac
      721 catacaccgg tgattaatcg tggtctacta ccaagcgtga gccacgtcgc cgacgaattt
      781 gagcagctct ggctgccgta ctggccgctg gcaagcgacg atctgctcga ggggatctac
      841 cgccaaagcc gcgcgtcggc cctaggccgc cggtacatcg aggcgaaccc aacagcgctg
      901 gcaaacctgc tggtcgtgga cgtagaccat ccagacgcag cgctccgagc gctcagcgcc
      961 cgggggtccc atccgctgcc caacgcgatc gtgggcaatc gcgccaacgg ccacgcacac
     1021 gcagtgtggg cactcaacgc ccctgttcca cgcaccgaat acgcgcggcg taagccgctc
     1081 gcatacatgg cggcgtgcgc cgaaggcctt cggcgcgccg tcgacggcga ccgcagttac
     1141 tcaggcctca tgaccaaaaa ccccggccac atcgcctggg aaacggaatg gctccactca
     1201 gatctctaca cactcagcca catcgaggcc gagctcggcg cgaacatgcc accgccgcgc
     1261 tggcgtcagc agaccacgta caaagcggct ccgacgccgc tagggcggaa ttgcgcactg
     1321 ttcgattccg tcaggttgtg ggcctatcgt cccgccctca tgcggatcta cctgccgacc
     1381 cggaacgtgg acggactcgg ccgcgcgatc tatgccgagt gccacgcgcg aaacgccgaa
     1441 ttcccgtgca acgacgtgtg tcccggaccg ctaccggaca gcgaggtccg cgccatcgcc
     1501 aacagcattt ggcgttggat cacaaccaag tcgcgcattt gggcggacgg gatcgtggtc
     1561 tacgaggcca cactcagtgc gcgccagtcg gccatctcgc ggaagggcgc agcagcgcgc
     1621 acggcggcga gcacagttgc gcggcgcgca aagtccgcgt cagccatgga ggcattgcta
     1681 tgagcgacgg ctacagcgac ggctacagcg acggctacaa ccggcagccg actgtccgca
     1741 aaaagcggcg cgtgaccgcc gccgaaggcg ctcgaatcac cggactatcc gaacgccacg
     1801 tcgtccggct cgtggcgcag gaacgcagcg agtggctcgc cgagcaggct gcacgccgcg
     1861 aacgcatccg cgcctatcac gacgacgagg gccactcttg gccgcaaacg gccaaacatt
     1921 tcgggctgca tctggacacc gttaagcgac tcggctatcg ggcgaggaaa gagcgtgcgg
     1981 cagaacagga agcggctcaa aaggcccaca acgaagccga caatccaccg ctgttctaac
     2041 gcaattgggg agcgggtgtc gcgggggttc cgtggggggt tccgttgcaa cgggtcggac
     2101 aggtaaaagt cctggtagac gctagttttc tggtttgggc catgcctgtc tcgttgcgtg
     2161 tttcgttgcg cccgttttga ataccagcca gacgagacgg ggttctacga atcttggtcg
     2221 ataccaagcc atttccgctg aatatcgggg agctcaccgc cagaatcggt ggttgtggtg
     2281 atgtacgtgg cgaactccgt tgtagtgcct gtggtggcat ccgtggccac tctcgttgca
     2341 cggttcgttg tgccgttaca ggccccgttg acagctcacc gaacgtagtt aaaacatgct
     2401 ggtcaaacta ggtttaccaa cgatacgagt cagctcatct agggccagtt ctaggcgttg
     2461 ttcgttgcgc ggttcgttgc gcatgtttcg tgtggttgct agatggctcc gcaaccacac
     2521 gcttcgaggt tgagtgcttc cagcacgggc gcgatccaga agaacttcgt cgtgcgactg
     2581 tcctcgttat ttctaggcac agcttcacgc tgccgcaagc actcagggcg caagggctgc
     2641 taaaggaagc ggaacacgta gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat
     2701 gtcagctact gggctatctg gacaagggaa aacgcaagcg caaagagaaa gcaggtagct
     2761 tgcagtgggc ttacatggcg atagctagac tgggcggttt tatggacagc aagcgaaccg
     2821 gaattgccag ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg
     2881 gctttcttgc cgccaaggat ctgatggcgc aggggatcaa gatctgatca agagacagga
     2941 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3001 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3061 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3121 gccctgaatg aactccaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3181 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3241 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3301 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3361 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3421 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3481 gcgcggatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3541 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3601 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3661 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3721 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     3781 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     3841 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     3901 tgctggagtt cttcgcccac cccctgtccc tcctgttcag ctactgacgg ggtggtgcgt
     3961 aacggcaaaa gcaccgccgg acatcagcgc cattcgccat tcaggctgcg caactgttgg
     4021 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     4081 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     4141 gccagtgcca agctgcgagc tcgaattctg attaacttta taaggaggaa aaacatatga
     4201 cagaagcttg gatccccggg aattcgtaat catggtcata gctgtttcct gtgtgaaatt
     4261 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4321 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4381 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4441 tgcgtattgg gcgctagcgg agtgtatata ctggcttact atgttggcac tgatgagggt
     4501 gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt cagcagaata
     4561 tgtgatacag gatatattcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac
     4621 tgcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat gccaggaaga
     4681 tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc tccgcccccc
     4741 tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata
     4801 aagataccag gcgtttcccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg
     4861 tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag
     4921 ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga
     4981 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc
     5041 accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg
     5101 gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc
     5161 ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt
     5221 cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta
     5281 atcagataaa at
//
