LOCUS       Exported                3420 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAA56, complete sequence.
ACCESSION   JF934877
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3420)
  AUTHORS   Mandavia CH, Figurski DH.
  TITLE     A new and better suicide vector for Aggregatibacter 
            actinomycetemcomitans
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3420)
  AUTHORS   Mandavia CH, Figurski DH.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-MAY-2011) Microbiology and Immunology, Columbia 
            University, 701 West 168th Street HHSC 1514A, New York, NY 10032, 
            USA
REFERENCE   3  (bases 1 to 3420)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3420
                     /organism="Cloning vector pAA56"
                     /mol_type="other DNA"
                     /db_xref="taxon:1094419"
     misc_feature    71..714
                     /note="Tra1 core containing the oriT; provides in trans 
                     oriT for mobilization of gene of interest by conjugation 
                     into Aggregatibacter actinomycetemcomitans"
     oriT            169..280
                     /rpt_unit_range=228 .. 267
                     /note="plasmid RK2 oriT sequence; potential for cruciform 
                     or hairpin formation"
     oriT            169..278
                     /note="incP origin of transfer"
     promoter        746..850
                     /gene="bla"
                     /label=AmpR promoter
     CDS             851..1711
                     /codon_start=1
                     /gene="bla"
                     /product="beta lactamase"
                     /label=bla
                     /note="penicillin resistance protein"
                     /protein_id="AEQ30016.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            851..1711
                     /gene="bla"
                     /label=bla
     gene            complement(1712..1779)
                     /gene="neo"
                     /label=neo
                     /note="residual kanamycin gene sequence left over from 
                     recombineering; derived from Tn5"
     misc_feature    1780..1795
                     /note="uptake signal sequence (USS) from tfox gene of 
                     Aggregatibacter aphrophilus; required for DNA uptake 
                     (transformation) into Aggregatibacter 
                     actinomycetemcomitans"
     rep_origin      1966..2554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2842..2863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2878..2908
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2916..2932
                     /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     2940..2956
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2952..3275
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    2969..3025
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3026..3042)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"