pMK228 vector (V016567)

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LOCUS       62056_17020        4588 bp DNA     circular SYN 27-JUL-2020
DEFINITION  Cloning vector pMK228, complete sequence.
ACCESSION   MN617162
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4588)
  AUTHORS   Hartsough LA, Kotlajich MV, Han B, Lin C-C.J., Gambill L, Wang MC, 
            Tabor JJ.
  TITLE     Optogenetic control of gut bacterial metabolism
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4588)
  AUTHORS   Hartsough LA, Kotlajich MV, Han B, Lin C-C.J., Gambill L, Wang MC, 
            Tabor JJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-OCT-2019) Bioengineering, Rice University, 6500 Main 
            St., Houston, TX 77030, USA
REFERENCE   3  (bases 1 to 4588)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-OCT-2019) Bioengineering, Rice University, 6500 Main St., 
            Houston, TX 77030, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      24..171
                     /note="specR promoter; from KD13"
                     /regulatory_class="promoter"
     CDS             172..960
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     terminator      975..1069
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1183..1728
                     /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."
     misc_feature    1984..2018
                     /label=constitutive promoter J23106
                     /note="constitutive promoter J23106"
     misc_feature    2019..2053
                     /label=B0034
                     /note="B0034"
     CDS             2054..4315
                     /codon_start=1
                     /transl_table=11
                     /product="CcaS"
                     /label=CcaS
                     /protein_id="QLL27773.1"
                     /translation="MGKFLIPIEFVFLAIAMTCYLWHRQNQERRRIEISIKQQTQRERF
                     INQITQHIRQSLNLETVLNTTVAEVKTLLQVDRVLIYRIWQDGTGSAITESVNANYPSI
                     LGRTFSDEVFPVEYHQAYTKGKVRAINDIDQDDIEICLADFVKQFGVKSKLVVPILQHN
                     RASSLDNESEFPYLWGLLITHQCAFTRPWQPWEVELMKQLANQVAIAIQQSELYEQLQQ
                     LNKDLENRVEKRTQQLAATNQSLRMEISERQKTEAALRHTNHTLQSLIAASPRGIFTLN
                     LADQIQIWNPTAERIFGWTETEIIAHPELLTSNILLEDYQQFKQKVLSGMVSPSLELKC
                     QKKDGSWIEIVLSAAPLLDSEENIAGLVAVVADITEQKRQAEQIRLLQSVVVNTNDAVV
                     ITEAEPIDDPGPRILYVNEAFTKITGYTAEEMLGKTPRVLQGPKTSRTELDRVRQAISQ
                     WQSVTVEVINYRKDGSEFWVEFSLVPVANKTGFYTHWIAVQRDVTERRRTEEVRLALER
                     EKELSRLKTRFFSMASHEFRTPLSTALAAAQLLENSEVAWLDPDKRSRNLHRIQNSVKN
                     MVQLLDDILIINRAEAGKLEFNPNWLDLKLLFQQFIEEIQLSVSDQYYFDFICSAQDTK
                     ALVDERLVRSILSNLLSNAIKYSPGGGQIKIALSLDSEQIIFEVTDQGIGISPEDQKQI
                     FEPFHRGKNVRNITGTGLGLMVAKKCVDLHSGSILLKSAVDQGTTVTICLKRYNHLPRA
                     "
     terminator      4336..4407
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4423..4450
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"