LOCUS       Exported                6778 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pMIND
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6778)
  AUTHORS   Blokpoel MC, Murphy HN, OToole R, Wiles S, Runn ES, Stewart GR, 
            Young DB, Robertson BD
  TITLE     Tetracycline-inducible gene regulation in mycobacteria.
  JOURNAL   Nucleic Acids Res. 2005  . 33(2):e22.
  PUBMED    15687380
REFERENCE   2  (bases 1 to 6778)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2005  . 33(2):e22."
FEATURES             Location/Qualifiers
     source          1..6778
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     19..35
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     19..35
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1916..1933)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1916..1932)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1925..1947)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4745..5539
                     /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(4799..4818)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5409..5428
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      5823..6411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6312..6331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6565..6582
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6699..6720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6735..6765
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(6773..6778,1..11)
                     /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     join(6778,1..22)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"