LOCUS       Exported               12273 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Constitutive trancription of FnCpf1 and recT in C.glutamitum, tac 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pJYS1Ptac
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12273)
  AUTHORS   Jiang Y, Qian F, Yang J, Liu Y, Dong F, Xu C, Sun B, Chen B, Xu X, 
            Li Y, Wang R, Yang S
  TITLE     CRISPR-Cpf1 assisted genome editing of Corynebacterium glutamicum.
  JOURNAL   Nat Commun. 2017 May 4;8:15179. doi: 10.1038/ncomms15179.
  PUBMED    28469274
REFERENCE   2  (bases 1 to 12273)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2017-05-4"; volume: "8:15179. doi"; pages: " 
            10.1038/ncomms15179"
FEATURES             Location/Qualifiers
     source          1..12273
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             3671..4465
                     /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(3725..3744)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4335..4354
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      8504..8531
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      8623..8708
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     9848..9867
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(9887..9964)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     promoter        10198..10226
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    10234..10250
                     /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     10239..10261
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             complement(11256..12206)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      join(12254..12273,1..203)
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"