LOCUS       Exported                6721 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTRKH2, complete sequence.
ACCESSION   MG417047
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6721)
  AUTHORS   Welker DL, Hughes JE, Steele JL, Broadbent JR.
  TITLE     High efficiency electrotransformation of Lactobacillus casei
  JOURNAL   FEMS Microbiol. Lett. 362 (2), 1-6 (2015)
  PUBMED    25670703
REFERENCE   2  (bases 1 to 6721)
  AUTHORS   Welker DL, Hughes JE, Steele JL, Broadbent JR.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-NOV-2017) Biology, Utah State University, 5305 Old 
            Main Hill, Logan, UT 84322-5305, USA
REFERENCE   3  (bases 1 to 6721)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6721
                     /organism="Cloning vector pTRKH2"
                     /mol_type="other DNA"
                     /db_xref="taxon:2315428"
     protein_bind    102..123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        138..168
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    176..192
                     /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     200..216
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             212..454
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITNSSISRSRARESLAAGRRIPGNSLAVVLQRRDWENPGVTQ
                     LNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEC"
     primer_bind     complement(295..311)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             992..1729
                     /codon_start=1
                     /gene="ermB"
                     /product="erythromycin resistance ErmB"
                     /label=ermB
                     /note="23S rRNA (adenine(2058)-N(6)-methyltransferase"
                     /protein_id="AXU41050.1"
                     /translation="MNKNIKYSQNFLTSEKVLNQIIKQLNLKETDTVYEIGTGKGHLTT
                     KLAKISKQVTSIELDSHLFNLSSEKLKLNTRVTLIHQDILQFQFPNKQRYKIVGSIPYH
                     LSTQIIKKVVFESHASDIYLIVEEGFYKRTLDIHRTLGLLLHTQVSIQQLLKLPAECFH
                     PKPKVNSVLIKLTRHTTDVPDKYWKLYTYFVSKWVNREYRQLFTKNQFHQAMKHAKVNN
                     LSTVTYEQVLSIFNSYLLFNGRK"
     gene            992..1729
                     /gene="ermB"
                     /label=ermB
     regulatory      2384..5298
                     /regulatory_class="replication_regulatory_region"
                     /note="Enterococcus origin region from pAMBeta1"
     CDS             3312..4802
                     /codon_start=1
                     /gene="repR"
                     /product="RepR"
                     /label=repR
                     /note="DNA replication protein"
                     /protein_id="AXU41049.1"
                     /translation="MNIPFVVETVLHDGLLKYKFKNSKIRSITTKPGKSKGAIFAYRSK
                     SSMIGGRGVVLTSEEAIQENQDTFTHWTPNVYRYGTYADENRSYTKGHSENNLRQINTF
                     FIDFDIHTAKETISASDILTTAIDLGFMPTMIIKSDKGYQAYFVLETPVYVTSKSEFKS
                     VKAAKIISQNIREYFGKSLPVDLTCNHFGIARIPRTDNVEFFDPNYRYSFKEWQDWSFK
                     QTDNKGFTRSSLTVLSGTEGKKQVDEPWFNLLLHETKFSGEKGLIGRNNVMFTLSLAYF
                     SSGYSIETCEYNMFEFNNRLDQPLEEKEVIKIVRSAYSENYQGANREYITILCKAWVSS
                     DLTSKDLFVRQGWFKFKKKRSERQRVHLSEWKEDLMAYISEKSDVYKPYLVTTKKEIRE
                     VLGIPERTLDKLLKVLKANQEIFFKIKPGRNGGIQLASVKSLLLSIIKVKKEEKESYIK
                     ALTNSFDLEHTFIQETLNKLAERPKTDTQLDLFSYDTG"
     gene            3312..4802
                     /gene="repR"
                     /label=repR
     promoter        complement(5512..5614)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     regulatory      6064..6564
                     /regulatory_class="replication_regulatory_region"
                     /note="Escherichia coli origin region from p15A"
     rep_origin      complement(6140..6685)
                     /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."