pAL36 vector (V009529)

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LOCUS       40924_4329       11418 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Reporter vector pAL36, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11418)
  AUTHORS   Karim AA, Gestaut DR, Fincker M, Ruth JC, Holmes EC, Sheu W, 
            Spormann AM.
  TITLE     Fine-tuned Protein Production in Methanosarcina acetivorans C2A
  JOURNAL   ACS Synth Biol (2018) In press
  PUBMED    29920209
REFERENCE   2  (bases 1 to 11418)
  AUTHORS   Karim AA.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-NOV-2017) Civil and Environmental Engineering, 
            Stanford University, Y2E2, 473 Via Ortega, Stanford, CA 94305, USA
REFERENCE   3  (bases 1 to 11418)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 11418)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol (2018) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-NOV-2017) Civil and Environmental Engineering, Stanford 
            University, Y2E2, 473 Via Ortega, Stanford, CA 94305, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      7..90
                     /label=minimal mcrB promoter with tetracycline operator
                     /note="minimal mcrB promoter with tetracycline operator"
                     /regulatory_class="promoter"
     protein_bind    46..63
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      91..109
                     /label=mcrB RBS from Methanosarcina acetivorans C2A
                     /note="mcrB RBS from Methanosarcina acetivorans C2A"
                     /regulatory_class="ribosome_binding_site"
     CDS             110..3181
                     /label=lacZ
                     /note="beta-galactosidase"
     misc_feature    3192..8662
                     /label=pC2A sequence
                     /note="pC2A sequence"
     CDS             complement(3798..4742)
                     /codon_start=1
                     /gene="ssrA"
                     /product="site-specific recombinase A"
                     /label=ssrA
                     /protein_id="AWW92034.1"
                     /translation="MGEMAAIQLTGIYEPLDNERLISLFTSDCIGRGYSKRTIEGYVSH
                     AKFFLQMCGVNAGFPELEGFLVHIRDERQYTLATCNNYFASLSTFFDFLEFKHIIERNN
                     IPTFRKRYLRSYKKYHTPETRQLITIEDMAKLVKVPRYPMYQALIIFLAKTGIRRNELI
                     TLDRADIDLEKCTVRLKKTAKRSNRIVFFDLETKAFLEAYLNTRHDKEKALFIGRQNGH
                     RVQRNLVYDIVTSAASSIGLHNPEGRLEEHFTPHCCRHWFTTWLRRSGMERTFIQELRG
                     DSRGEAIDVYDHIEKDELREAYLKCIPQLGIKP"
     gene            complement(3798..4742)
                     /gene="ssrA"
                     /label=ssrA
     misc_feature    5225..5872
                     /label=Orf2
                     /note="Orf2"
     misc_feature    complement(5878..6306)
                     /label=Orf1
                     /note="Orf1"
     CDS             complement(6385..7995)
                     /codon_start=1
                     /gene="repA"
                     /product="regulatory protein RepA"
                     /label=repA
                     /protein_id="AWW92035.1"
                     /translation="MSSDFSSRPGAVSYAPECRDFATCICTDGVLCPVRNKKNGVSICK
                     YYSCPDFDFSKCSKYRPLKVGDFVLPAKGKRPAYCGRPFTYAVSENYNARKDLNSHCNT
                     LSCPGCGILSELQHRFALVVILWTYALFSGQFPSWGFASMDFSRGVSVEAIRLFRRNLK
                     DRLERWGVTAGTTIFHPYRIKPEITRAIRKLTGLKGNDDSEAFWNYIRDDHNEGNLNKI
                     ADLLNIEINSIYDCLILAPHFHFFIFPGNIRITGKNPKSKKRCKSDEYDIFIQRLSHKV
                     NGKRQWELRSLMDIYKQVTYLYSHVGQLTDNRYGNIHIESRFGALFRWHPDKVVSSSGK
                     LTFEQLIDTRTEVLNLINEGREVKLVYNGELKWEGQENDDTGWIPLTKLRDHFSGGRQA
                     LHAYLKEAKAVKPEYADYLQQLIARNNFYHNLKGRDGKNKVPKKWRKLWASPFKVDELP
                     EFLKLEPAGSLLRVWFLEGLPDPPAGFLVVDSHQWEHDGCIIEESPEMPSNSLQKASEI
                     NYEELFTSLKSRSMMFGGD"
     gene            complement(6385..7995)
                     /gene="repA"
                     /label=repA
     regulatory      8659..9020
                     /label=mcr promoter from Methanococcus voltae
                     /note="mcr promoter from Methanococcus voltae"
                     /regulatory_class="promoter"
     CDS             9021..9617
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     regulatory      9621..9960
                     /label=mcr terminator from Methanococcus voltae
                     /note="mcr terminator from Methanococcus voltae"
                     /regulatory_class="terminator"
     rep_origin      complement(9978..10366)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     regulatory      complement(10395..10427)
                     /label=bla terminator
                     /note="bla terminator"
                     /regulatory_class="terminator"
     CDS             complement(10431..11288)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(11289..11393)
                     /label=AmpR promoter