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  JOURNAL   Unpublished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            "Unpublished"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"
ORIGIN
        1 tcgacctgca ggcatgcaag cttagcaagc gaaccggaat tgccagctgg ggcgccctct
       61 ggtaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc aaggatctga
      121 tggcgcaggg gatcaagatc tgatcaagag acaggatgag gatcgtttcg catgagggaa
      181 gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat
      241 ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag
      301 ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg
      361 cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc
      421 cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct
      481 aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag
      541 ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt
      601 gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt
      661 gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag
      721 cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg
      781 ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc
      841 atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca
      901 gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa
      961 taagagctcg cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga
     1021 tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc caagcactag
     1081 taacaactta tatcgtatgg ggctgacttc aggtgctaca tttgaagaga taaattgcac
     1141 tgaaatctag aaatatttta tctgattaat aagatgatct tcttgagatc gttttggtct
     1201 gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt
     1261 tctctgagct accaactctt tgaaccgagg taactggctt ggaggagcgc agtcaccaaa
     1321 acttgtcctt tcagtttagc cttaaccggc gcatgacttc aagactaact cctctaaatc
     1381 aattaccagt ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg gactcaagac
     1441 gatagttacc ggataaggcg cagcggtcgg actgaacggg gggttcgtgc atacagtcca
     1501 gcttggagcg aactgcctac ccggaactga gtgtcaggcg tggaatgaga caaacgcggc
     1561 cataacagcg gaatgacacc ggtaaaccga aaggcaggaa caggagagcg cacgagggag
     1621 ccgccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca ccactgattt
     1681 gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa cggctttgcc
     1741 gcggccctct cacttccctg ttaagtatct tcctggcatc ttccaggaaa tctccgcccc
     1801 gttcgtaagc catttccgct cgccgcagtc gaacgaccga gcgtagcgag tcagtgagcg
     1861 aggaagcgga atatatcccc taggtggagt ctggcctcaa atacaaagtc tgcaaagtct
     1921 cgaaatatta aaccagagaa taaataatta aacgccaaaa tatttctagc aaaaacccca
     1981 gtttttacgg ctagctcagt cctaggtata gtgctagcta tagaggttaa aatcttaata
     2041 ggaggaaggc gcaatgggca aatttctaat tccaatcgaa tttgtttttc tggcgatcgc
     2101 catgacctgt tatttatggc acagacaaaa ccaagaacgc cgcaggattg aaattagcat
     2161 caagcaacaa acccaacggg aacgatttat taaccaaatt acccaacata tccgccaatc
     2221 tttaaacttg gaaacggttt taaataccac cgtcgctgaa gttaaaaccc tgttgcaagt
     2281 tgatcgagtt ctaatttatc gcatttggca agatggcacg ggcagcgcca ttacggaatc
     2341 ggtgaatgcc aattatccta gtattttagg gcggaccttt tccgatgaag tttttcccgt
     2401 tgaataccat caagcctaca ccaaaggtaa agtacgggcc attaatgaca ttgaccagga
     2461 tgacatagag atttgcctag ctgatttcgt caaacaattt ggcgtgaaat caaaattagt
     2521 agtgcccatt cttcaacata atcgtgcttc ttccctagat aatgaatcag aatttcccta
     2581 tctttggggg ctgttaatta cccatcaatg tgcttttacc cggccatggc aaccgtggga
     2641 agtggagtta atgaaacagc tagccaatca ggtcgcgatc gccatccaac aatcggaatt
     2701 atatgagcaa ttacagcaac tcaataaaga tttggaaaac cgagtcgaaa aacgcaccca
     2761 gcaacttgcc gccaccaatc aatccctaag aatggaaatc agtgagcgac aaaaaacgga
     2821 agccgctctc cgccacacta accatactct gcaatccctg attgcggcct cccccagggg
     2881 tatttttacc cttaatttag cagaccaaat tcagatttgg aatcctacag cagaacgtat
     2941 ttttggttgg acagaaacag aaattattgc ccatccagaa ttattaacat ccaacatttt
     3001 gctggaagat tatcagcaat ttaaacagaa agttttatca ggcatggttt cccctagcct
     3061 agaattaaaa tgtcaaaaaa aagatggtag ttggattgaa attgtccttt ccgctgctcc
     3121 cctattggat agtgaagaaa atattgccgg attggtggcg gttgtcgccg atattaccga
     3181 gcaaaagcgg caggcagaac aaattcgttt gctacaatcc gttgtggtta atactaatga
     3241 tgcggtggtg attacggaag cggagcccat tgatgatccc gggccgagaa ttctctatgt
     3301 caatgaagca tttactaaaa tcaccggtta tactgctgaa gaaatgctag gcaaaacccc
     3361 ccgagtttta cagggaccaa aaactagtcg cactgaatta gatagggtgc ggcaagccat
     3421 tagtcaatgg caatcagtta ccgttgaagt gattaattat cgtaaggatg gcagtgagtt
     3481 ttgggtggaa tttagtctgg tgcccgttgc caataaaaca ggtttttaca cccattggat
     3541 tgctgtgcaa agggatgtca ctgagcgccg acgcacggag gaagtccgcc tagctttaga
     3601 acgggaaaaa gaattaagcc gcctaaaaac tcgttttttc tccatggctt cccatgaatt
     3661 tcgtactccc ctcagtacgg ccttagctgc tgcccaatta ctggaaaatt ctgaagtggc
     3721 ctggcttgat cccgataagc gtagccggaa cttacaccgt attcaaaatt ccgtgaaaaa
     3781 tatggtacag ctcctggatg atattttaat cattaaccgt gccgaagcgg gcaaattgga
     3841 atttaatcct aattggttag atttgaaatt attgttccag caatttatcg aagaaattca
     3901 attaagtgtc agtgaccaat attattttga ctttatttgt agcgctcaag atacgaaggc
     3961 attggtggat gaaaggttag tgcggtctat tttatctaat ctgttatcta atgcgattaa
     4021 atactctccc gggggagggc agattaaaat tgccctaagc ctagattcgg aacagattat
     4081 ttttgaagtc accgaccagg gcattggcat ttcgccagag gaccaaaagc aaatttttga
     4141 accctttcat cggggcaaaa atgtcagaaa tattacggga acaggactcg gtttaatggt
     4201 tgccaagaaa tgtgttgact tacacagtgg cagtatcttg ctaaaaagtg cagttgacca
     4261 gggaacaaca gttactatct gtttaaaacg ctataaccat ttgcctcgag cttagtaata
     4321 atctagacca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     4381 atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg
     4441 cctttctgcg tttatagtaa atcactgcat aattcgtgtc gattaaaccg cctaagtccc
     4501 ccaggaaagg gggatataac agtatagatt ttgtcagcct tcagcttggc tttaccgtca
     4561 aaaaaaggta cccggggatc ctctagag
//
