LOCUS       sequence-222515-        5317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle vector for high level expression 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: pSUM36-MCS2-myc
FEATURES             Location/Qualifiers
     source          1..5317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     39..56
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    293..314
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(329..334,335..352,353..359)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    367..383
                     /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     372..394
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     391..407
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     391..407
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             403..693
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITNSRGSSQIFFRNKFLFQSFCHMFFLLIKLIRIRARSLALA
                     VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWR"
     CDS             complement(436..465)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(531..548)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(531..547)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(540..562)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(927..1721)
                     /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(1038..1057)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1648..1667
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      4694..5239
                     /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     5140..5159
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aagccatttc cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag
       61 cggaatatat cctgtatcac atattctgct gacgcaccgg tgcagccttt tttctcctgc
      121 cacatgaagc acttcactga caccctcatc agtgccaaca tagtaagcca gtatatacac
      181 tccgctagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
      241 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
      301 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
      361 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgaat
      421 tcccggggat cctcacagat cttcttcaga aataagtttt tgttccaaag cttctgtcat
      481 atgtttttcc tccttataaa gttaatcaga attcgagctc gcagcttggc actggccgtc
      541 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
      601 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
      661 cagttgcgca gcctgaatgg cgaatggcgc tgatgtccgg cggtgctttt gccgttacgc
      721 accaccccgt cagtagctga acaggaggga cagggggtgg gcgaagaact ccagcatgag
      781 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
      841 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc
      901 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg
      961 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
     1021 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
     1081 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
     1141 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat ccgcgccttg
     1201 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     1261 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     1321 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     1381 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     1441 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     1501 cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt ggagttcatt cagggcaccg
     1561 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     1621 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     1681 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct
     1741 gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc
     1801 cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt
     1861 tcgcttgctg tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct
     1921 acctgctttc tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc
     1981 atccggggtc agcaccgttt ctgcggactg gctttctacg tgttccgctt cctttagcag
     2041 cccttgcgcc ctgagtgctt gcggcagcgt gaagctgtgc ctagaaataa cgaggacagt
     2101 cgcacgacga agttcttctg gatcgcgccc gtgctggaag cactcaacct cgaagcgtgt
     2161 ggttgcggag ccatctagca accacacgaa acatgcgcaa cgaaccgcgc aacgaacaac
     2221 gcctagaact ggccctagat gagctgactc gtatcgttgg taaacctagt ttgaccagca
     2281 tgttttaact acgttcggtg agctgtcaac ggggcctgta acggcacaac gaaccgtgca
     2341 acgagagtgg ccacggatgc caccacaggc actacaacgg agttcgccac gtacatcacc
     2401 acaaccaccg attctggcgg tgagctcccc gatattcagc ggaaatggct tggtatcgac
     2461 caagattcgt agaaccccgt ctcgtctggc tggtattcaa aacgggcgca acgaaacacg
     2521 caacgagaca ggcatggccc aaaccagaaa actagcgtct accaggactt ttacctgtcc
     2581 gacccgttgc aacggaaccc cccacggaac ccccgcgaca cccgctcccc aattgcgtta
     2641 gaacagcggt ggattgtcgg cttcgttgtg ggccttttga gccgcttcct gttctgccgc
     2701 acgctctttc ctcgcccgat agccgagtcg cttaacggtg tccagatgca gcccgaaatg
     2761 tttggccgtt tgcggccaag agtggccctc gtcgtcgtga taggcgcgga tgcgttcgcg
     2821 gcgtgcagcc tgctcggcga gccactcgct gcgttcctgc gccacgagcc ggacgacgtg
     2881 gcgttcggat agtccggtga ttcgagcgcc ttcggcggcg gtcacgcgcc gctttttgcg
     2941 gacagtcggc tgccggttgt agccgtcgct gtagccgtcg ctcatagcaa tgcctccatg
     3001 gctgacgcgg actttgcgcg ccgcgcaact gtgctcgccg ccgtgcgcgc tgctgcgccc
     3061 ttccgcgaga tggccgactg gcgcgcactg agtgtggcct cgtagaccac gatcccgtcc
     3121 gcccaaatgc gcgacttggt tgtgatccaa cgccaaatgc tgttggcgat ggcgcggacc
     3181 tcgctgtccg gtagcggtcc gggacacacg tcgttgcacg ggaattcggc gtttcgcgcg
     3241 tggcactcgg catagatcgc gcggccgagt ccgtccacgt tccgggtcgg caggtagatc
     3301 cgcatgaggg cgggacgata ggcccacaac ctgacggaat cgaacagtgc gcaattccgc
     3361 cctagcggcg tcggagccgc tttgtacgtg gtctgctgac gccagcgcgg cggtggcatg
     3421 ttcgcgccga gctcggcctc gatgtggctg agtgtgtaga gatctgagtg gagccattcc
     3481 gtttcccagg cgatgtggcc ggggtttttg gtcatgaggc ctgagtaact gcggtcgccg
     3541 tcgacggcgc gccgaaggcc ttcggcgcac gccgccatgt atgcgagcgg cttacgccgc
     3601 gcgtattcgg tgcgtggaac aggggcgttg agtgcccaca ctgcgtgtgc gtggccgttg
     3661 gcgcgattgc ccacgatcgc gttgggcagc ggatgggacc cccgggcgct gagcgctcgg
     3721 agcgctgcgt ctggatggtc tacgtccacg accagcaggt ttgccagcgc tgttgggttc
     3781 gcctcgatgt accggcggcc tagggccgac gcgcggcttt ggcggtagat cccctcgagc
     3841 agatcgtcgc ttgccagcgg ccagtacggc agccagagct gctcaaattc gtcggcgacg
     3901 tggctcacgc ttggtagtag accacgatta atcaccggtg tatggtccga cacgagctcc
     3961 aagtcagata tttcgctgag gggccacccc acaactgcac actcccccgc tctcccgtcg
     4021 agccctggtg gtggaacacc agcgacagcc gagcaccccc aaccacctgt accaaccagg
     4081 aggaacacat gcgtcgtttc gaggacgttt ccgggccgct gagagccgct gtggcggccg
     4141 tacacgccgc cttagacccg ttagaccccc tgccgcctga atgcgcgggt acgagccaca
     4201 cagcgcccga acttacggag ctggtgggct cacctggctt tatggcgtac gaatcggctg
     4261 tgtgcgacct gttgggcgag gtgaggtacg cgctactcac gctggcaagg gcgacagagc
     4321 cgccccaccg agcccgcacg gccgcgcgcg gtgtcaacaa ccgggtgagt cgtgcacacc
     4381 agcaggtgtt cgaggcttgg ctcgaagtgc aggacatcgt ggcgaacgcc gcccgatgag
     4441 ccgcgcctta cgctggctgc cagccgttcg cgggctggtt ggtgcagcgc gtcgagcggt
     4501 tagaggccct gcggtgttcc accaccgcag ggcctcgccc tttttaaggc tgaatttgct
     4561 tgtctccgaa tccaactggc ttgtccaagg gtgtatctac gcttaatcca aagttcaaac
     4621 gaggggatta cacatgacca acttcgataa cgttctcggc tcgatatttt atctgattaa
     4681 taagatgatc ttcttgagat cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa
     4741 aaaccgcctt gcagggcggt ttttcgaagg ttctctgagc taccaactct ttgaaccgag
     4801 gtaactggct tggaggagcg cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg
     4861 cgcatgactt caagactaac tcctctaaat caattaccag tggctgctgc cagtggtgct
     4921 tttgcatgtc tttccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4981 gactgaacgg ggggttcgtg catacagtcc agcttggagc gaactgccta cccggaactg
     5041 agtgtcaggc gtggaatgag acaaacgcgg ccataacagc ggaatgacac cggtaaaccg
     5101 aaaggcagga acaggagagc gcacgaggga gccgccaggg ggaaacgcct ggtatcttta
     5161 tagtcctgtc gggtttcgcc accactgatt tgagcgtcag atttcgtgat gcttgtcagg
     5221 ggggcggagc ctatggaaaa acggctttgc cgcggccctc tcacttccct gttaagtatc
     5281 ttcctggcat cttccaggaa atctccgccc cgttcgt
//
