LOCUS       40924_2997        3420 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAA56, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3420)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-MAY-2011) Microbiology and Immunology, Columbia University, 701 
            West 168th Street HHSC 1514A, New York, NY 10032, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            complement(169..278)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        746..850
                     /label=AmpR promoter
     CDS             851..1708
                     /label=AmpR
                     /note="beta-lactamase"
     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
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2842..2863
                     /label=CAP binding site
                     /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
                     /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"
     misc_feature    complement(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"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc gtctggaagg cagtacacct tgataggtgg gctgcccttc ctggttggct
      121 tggtttcatc agccatccgc ttgccctcat ctgttacgcc ggcggtagcc ggccagcctc
      181 gcagagcagg attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac
      241 acccgctcgc gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc
      301 aaggaaagtc tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat
      361 ataccgaaaa aatcgctata atgaccccga agcagggtta tgcagcggaa aagcgctgct
      421 tccctgctgt tttgtggaat atctaccgac tggaaacagg caaatgcagg aaattactga
      481 actgagggga caggcgagag acgatgccaa agagctacac cgacgagctg gccgagtggg
      541 ttgaatcccg cgcggccaag aagcgccggc gtgatgaggc tgcggttgcg ttcctggcgg
      601 tgagggcgga tgtcgaggcg gcgttagcgt ccggctatgc gctcgtcacc atttgggagc
      661 acatgcggga aacggggaag gtcaagttct cctacgagac gttccgctcg cacggacgtc
      721 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
      781 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      841 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      901 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      961 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1021 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1081 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     1141 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     1201 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1261 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     1321 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     1381 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     1441 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     1501 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     1561 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     1621 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     1681 gataggtgcc tcactgatta agcattggta agatgctcga tgagtttttc taatcagaat
     1741 tggttaattg gttgtaacac tggcagagca ttacgaaata aagtgcggtc ggttcctgtc
     1801 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     1861 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     1921 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1981 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2041 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2101 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2161 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2221 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2281 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2341 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2401 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2461 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2521 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2581 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2641 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2701 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     2761 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     2821 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     2881 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     2941 aggaaacagc tatgaccatg attacgccaa gcttgcatgc ctgcaggtcg actctagagg
     3001 atccccgggt accgagctcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     3061 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     3121 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     3181 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     3241 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     3301 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3361 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
//
