LOCUS       sequence-211589-        5317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses proteins in mycobacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5317)
  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 5317)
  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-MCS3
FEATURES             Location/Qualifiers
     source          1..5317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..291
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITNSRGSDIHVFPPYKVNQSFCHMFFLLIKLIRIRARSLALA
                     VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWR"
     primer_bind     complement(129..146)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(129..145)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(138..160)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(525..1319)
                     /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(636..655)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1246..1265
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      4292..4837
                     /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     4738..4757
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4954..4971
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5208..5229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5244..5249,5250..5267,5268..5274)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5282..5298
                     /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     5287..5309
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(5306..5317,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5306..5317,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgaattc ccggggatcc gatatccatg tttttcctcc ttataaagtt
       61 aatcaaagct tctgtcatat gtttttcctc cttataaagt taatcagaat tcgagctcgc
      121 agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
      181 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
      241 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctg atgtccggcg
      301 gtgcttttgc cgttacgcac caccccgtca gtagctgaac aggagggaca gggggtgggc
      361 gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac
      421 gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag
      481 gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca
      541 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
      601 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
      661 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
      721 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
      781 tcgggcatcc gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
      841 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
      901 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
      961 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     1021 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     1081 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgg
     1141 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     1201 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     1261 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
     1321 cgaaacgatc ctcatcctgt ctcttgatca gatcttgatc ccctgcgcca tcagatcctt
     1381 ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc
     1441 ccagctggca attccggttc gcttgctgtc cataaaaccg cccagtctag ctatcgccat
     1501 gtaagcccac tgcaagctac ctgctttctc tttgcgcttg cgttttccct tgtccagata
     1561 gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc tttctacgtg
     1621 ttccgcttcc tttagcagcc cttgcgccct gagtgcttgc ggcagcgtga agctgtgcct
     1681 agaaataacg aggacagtcg cacgacgaag ttcttctgga tcgcgcccgt gctggaagca
     1741 ctcaacctcg aagcgtgtgg ttgcggagcc atctagcaac cacacgaaac atgcgcaacg
     1801 aaccgcgcaa cgaacaacgc ctagaactgg ccctagatga gctgactcgt atcgttggta
     1861 aacctagttt gaccagcatg ttttaactac gttcggtgag ctgtcaacgg ggcctgtaac
     1921 ggcacaacga accgtgcaac gagagtggcc acggatgcca ccacaggcac tacaacggag
     1981 ttcgccacgt acatcaccac aaccaccgat tctggcggtg agctccccga tattcagcgg
     2041 aaatggcttg gtatcgacca agattcgtag aaccccgtct cgtctggctg gtattcaaaa
     2101 cgggcgcaac gaaacacgca acgagacagg catggcccaa accagaaaac tagcgtctac
     2161 caggactttt acctgtccga cccgttgcaa cggaaccccc cacggaaccc ccgcgacacc
     2221 cgctccccaa ttgcgttaga acagcggtgg attgtcggct tcgttgtggg ccttttgagc
     2281 cgcttcctgt tctgccgcac gctctttcct cgcccgatag ccgagtcgct taacggtgtc
     2341 cagatgcagc ccgaaatgtt tggccgtttg cggccaagag tggccctcgt cgtcgtgata
     2401 ggcgcggatg cgttcgcggc gtgcagcctg ctcggcgagc cactcgctgc gttcctgcgc
     2461 cacgagccgg acgacgtggc gttcggatag tccggtgatt cgagcgcctt cggcggcggt
     2521 cacgcgccgc tttttgcgga cagtcggctg ccggttgtag ccgtcgctgt agccgtcgct
     2581 catagcaatg cctccatggc tgacgcggac tttgcgcgcc gcgcaactgt gctcgccgcc
     2641 gtgcgcgctg ctgcgccctt ccgcgagatg gccgactggc gcgcactgag tgtggcctcg
     2701 tagaccacga tcccgtccgc ccaaatgcgc gacttggttg tgatccaacg ccaaatgctg
     2761 ttggcgatgg cgcggacctc gctgtccggt agcggtccgg gacacacgtc gttgcacggg
     2821 aattcggcgt ttcgcgcgtg gcactcggca tagatcgcgc ggccgagtcc gtccacgttc
     2881 cgggtcggca ggtagatccg catgagggcg ggacgatagg cccacaacct gacggaatcg
     2941 aacagtgcgc aattccgccc tagcggcgtc ggagccgctt tgtacgtggt ctgctgacgc
     3001 cagcgcggcg gtggcatgtt cgcgccgagc tcggcctcga tgtggctgag tgtgtagaga
     3061 tctgagtgga gccattccgt ttcccaggcg atgtggccgg ggtttttggt catgaggcct
     3121 gagtaactgc ggtcgccgtc gacggcgcgc cgaaggcctt cggcgcacgc cgccatgtat
     3181 gcgagcggct tacgccgcgc gtattcggtg cgtggaacag gggcgttgag tgcccacact
     3241 gcgtgtgcgt ggccgttggc gcgattgccc acgatcgcgt tgggcagcgg atgggacccc
     3301 cgggcgctga gcgctcggag cgctgcgtct ggatggtcta cgtccacgac cagcaggttt
     3361 gccagcgctg ttgggttcgc ctcgatgtac cggcggccta gggccgacgc gcggctttgg
     3421 cggtagatcc cctcgagcag atcgtcgctt gccagcggcc agtacggcag ccagagctgc
     3481 tcaaattcgt cggcgacgtg gctcacgctt ggtagtagac cacgattaat caccggtgta
     3541 tggtccgaca cgagctccaa gtcagatatt tcgctgaggg gccaccccac aactgcacac
     3601 tcccccgctc tcccgtcgag ccctggtggt ggaacaccag cgacagccga gcacccccaa
     3661 ccacctgtac caaccaggag gaacacatgc gtcgtttcga ggacgtttcc gggccgctga
     3721 gagccgctgt ggcggccgta cacgccgcct tagacccgtt agaccccctg ccgcctgaat
     3781 gcgcgggtac gagccacaca gcgcccgaac ttacggagct ggtgggctca cctggcttta
     3841 tggcgtacga atcggctgtg tgcgacctgt tgggcgaggt gaggtacgcg ctactcacgc
     3901 tggcaagggc gacagagccg ccccaccgag cccgcacggc cgcgcgcggt gtcaacaacc
     3961 gggtgagtcg tgcacaccag caggtgttcg aggcttggct cgaagtgcag gacatcgtgg
     4021 cgaacgccgc ccgatgagcc gcgccttacg ctggctgcca gccgttcgcg ggctggttgg
     4081 tgcagcgcgt cgagcggtta gaggccctgc ggtgttccac caccgcaggg cctcgccctt
     4141 tttaaggctg aatttgcttg tctccgaatc caactggctt gtccaagggt gtatctacgc
     4201 ttaatccaaa gttcaaacga ggggattaca catgaccaac ttcgataacg ttctcggctc
     4261 gatattttat ctgattaata agatgatctt cttgagatcg ttttggtctg cgcgtaatct
     4321 cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt ctctgagcta
     4381 ccaactcttt gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa cttgtccttt
     4441 cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca attaccagtg
     4501 gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg atagttaccg
     4561 gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag cttggagcga
     4621 actgcctacc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg
     4681 aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc cgccaggggg
     4741 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac cactgatttg agcgtcagat
     4801 ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac ggctttgccg cggccctctc
     4861 acttccctgt taagtatctt cctggcatct tccaggaaat ctccgccccg ttcgtaagcc
     4921 atttccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa
     4981 tatatcctgt atcacatatt ctgctgacgc accggtgcag ccttttttct cctgccacat
     5041 gaagcacttc actgacaccc tcatcagtgc caacatagta agccagtata tacactccgc
     5101 tagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     5161 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     5221 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     5281 attgtgagcg gataacaatt tcacacagga aacagct
//
