pAL23 vector (V009530)

Basic Vector Information

Vector Name:
pAL23
Antibiotic Resistance:
Ampicillin
Length:
8325 bp
Type:
Cloning vector
Replication origin:
R6K γ ori
Source/Author:
Karim AA, Gestaut DR, Fincker M, Ruth JC, Holmes EC, Sheu W, Spormann AM.

pAL23 vector Map

pAL238325 bp400800120016002000240028003200360040004400480052005600600064006800720076008000M13 fwdpC2A sequencePuroRmcr terminator from Methanococcus voltaeR6K gamma oribla terminatorAmpRAmpR promoter

pAL23 vector Sequence

LOCUS       40924_4324        8325 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAL23, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8325)
  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 8325)
  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 8325)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8325)
  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..8325
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     26..42
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    99..5569
                     /label=pC2A sequence
                     /note="pC2A sequence"
     CDS             complement(705..1649)
                     /codon_start=1
                     /gene="ssrA"
                     /product="site-specific recombinase A"
                     /label=ssrA
                     /protein_id="AWW92029.1"
                     /translation="MGEMAAIQLTGIYEPLDNERLISLFTSDCIGRGYSKRTIEGYVSH
                     AKFFLQMCGVNAGFPELEGFLVHIRDERQYTLATCNNYFASLSTFFDFLEFKHIIERNN
                     IPTFRKRYLRSYKKYHTPETRQLITIEDMAKLVKVPRYPMYQALIIFLAKTGIRRNELI
                     TLDRADIDLEKCTVRLKKTAKRSNRIVFFDLETKAFLEAYLNTRHDKEKALFIGRQNGH
                     RVQRNLVYDIVTSAASSIGLHNPEGRLEEHFTPHCCRHWFTTWLRRSGMERTFIQELRG
                     DSRGEAIDVYDHIEKDELREAYLKCIPQLGIKP"
     gene            complement(705..1649)
                     /gene="ssrA"
                     /label=ssrA
     misc_feature    2132..2779
                     /label=Orf2
                     /note="Orf2"
     misc_feature    complement(2785..3213)
                     /label=Orf1
                     /note="Orf1"
     CDS             complement(3292..4902)
                     /codon_start=1
                     /gene="repA"
                     /product="regulatory protein RepA"
                     /label=repA
                     /protein_id="AWW92030.1"
                     /translation="MSSDFSSRPGAVSYAPECRDFATCICTDGVLCPVRNKKNGVSICK
                     YYSCPDFDFSKCSKYRPLKVGDFVLPAKGKRPAYCGRPFTYAVSENYNARKDLNSHCNT
                     LSCPGCGILSELQHRFALVVILWTYALFSGQFPSWGFASMDFSRGVSVEAIRLFRRNLK
                     DRLERWGVTAGTTIFHPYRIKPEITRAIRKLTGLKGNDDSEAFWNYIRDDHNEGNLNKI
                     ADLLNIEINSIYDCLILAPHFHFFIFPGNIRITGKNPKSKKRCKSDEYDIFIQRLSHKV
                     NGKRQWELRSLMDIYKQVTYLYSHVGQLTDNRYGNIHIESRFGALFRWHPDKVVSSSGK
                     LTFEQLIDTRTEVLNLINEGREVKLVYNGELKWEGQENDDTGWIPLTKLRDHFSGGRQA
                     LHAYLKEAKAVKPEYADYLQQLIARNNFYHNLKGRDGKNKVPKKWRKLWASPFKVDELP
                     EFLKLEPAGSLLRVWFLEGLPDPPAGFLVVDSHQWEHDGCIIEESPEMPSNSLQKASEI
                     NYEELFTSLKSRSMMFGGD"
     gene            complement(3292..4902)
                     /gene="repA"
                     /label=repA
     regulatory      5566..5927
                     /label=mcr promoter from Methanococcus voltae
                     /note="mcr promoter from Methanococcus voltae"
                     /regulatory_class="promoter"
     CDS             5928..6524
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     regulatory      6528..6867
                     /label=mcr terminator from Methanococcus voltae
                     /note="mcr terminator from Methanococcus voltae"
                     /regulatory_class="terminator"
     rep_origin      complement(6885..7273)
                     /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(7302..7334)
                     /label=bla terminator
                     /note="bla terminator"
                     /regulatory_class="terminator"
     CDS             complement(7338..8195)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8196..8300)
                     /label=AmpR promoter

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