LOCUS       62056_8210        3844 bp DNA     circular SYN 14-NOV-2020
DEFINITION  Cloning vector pD1, complete sequence.
ACCESSION   MT180325
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3844)
  AUTHORS   Luo M, Xie L, Chakraborty S, Wang A, Matny O, Richardson T, Bhatt D,
            Hoque M, Patpour M, Sorenson C, Steuernagel B, Wulff B, Upadhyaya N,
            Mago R, Periyannan S, Lagudah E, Freedman R, Reuber L, Steffenson B,
            Ayliffe M.
  TITLE     A wheat stem rust resistance multi-transgene cassette provides high 
            levels of field protection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3844)
  AUTHORS   Luo M, Xie L, Chakraborty S, Wang A, Matny O, Richardson T, Bhatt D,
            Hoque M, Patpour M, Sorenson C, Steuernagel B, Wulff B, Upadhyaya N,
            Mago R, Periyannan S, Lagudah E, Freedman R, Reuber L, Steffenson B,
            Ayliffe M.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-MAR-2020) RP1, CSIRO A
REFERENCE   3  (bases 1 to 3844)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-MAR-2020) RP1, CSIRO A"
FEATURES             Location/Qualifiers
     source          1..3844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    33..132
                     /label=attL1
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    259..280
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        295..325
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    333..349
                     /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     357..373
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(398..414)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    671..902
                     /label=attP1
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    904..940
                     /label=PI-Sce I
                     /note="PI-Sce I"
     promoter        complement(944..962)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1226..1330
                     /label=AmpR promoter
     CDS             1331..2188
                     /label=AmpR
                     /note="beta-lactamase"
     protein_bind    complement(2287..2320)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(2354..2477)
                     /label=attR3
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    2479..2598
                     /label=multiple cloning site
                     /note="multiple cloning site"
     protein_bind    complement(2599..2698)
                     /label=attL2
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    2700..2717
                     /label=I-Sce I
                     /note="I-Sce I"
     rep_origin      2795..3383
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(3569..3709)
                     /label=bom
                     /note="basis of mobility region from pBR322"
ORIGIN
        1 gctgtagaag tcgtcatagc ttaccgaaat cgcaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg ctttggcaaa cagctattat gggtcgccca atacgcaaac cgcctctccc
      181 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      241 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
      301 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
      361 aaacagctat gaccatgatt acgccagata tcatgtcact ggccgtcgtt ttacaacgtc
      421 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
      481 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
      541 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
      601 cgcgcagcta agactatcat ttaggtgaca ctatagtgct acagtcagtt gctagacatc
      661 gactatgctc aaataatgat tttattttga ctgatagtga cctgttcgtt gcaacaaatt
      721 gatgagcaat gcttttttat aatgccaact ttgtacaaaa aagctgaacg agaaacgtaa
      781 aatgatataa atatcaatat attaaattag attttgcata aaaaacagac tacataatac
      841 tgtaaaacac aacatatcca gtcactatga atcaactact tagatggtat tagtgacctg
      901 tagccatttc attacctctt tctccgcacc cgacatagat ggccctatag tgagtcgtat
      961 taggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     1021 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     1081 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     1141 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     1201 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1261 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1321 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1381 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1441 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1501 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1561 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     1621 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     1681 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1741 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     1801 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     1861 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     1921 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     1981 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2041 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2101 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2161 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2221 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2281 taatctataa cttcgtatag catacattat acgaagttat tacaggtcac taataccatc
     2341 taagtagttg attcatagtg actgcatatg ttgtgtttta cagtattatg tagtctgttt
     2401 tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt ctcgttcaac
     2461 tttattatac aaagttgttg catgcggccg ctagctcgag aggcgcgcct accggtgagc
     2521 tctagaattc tgcaggtacc gcggatccat gggcccggga ctagtcgaca tgtacaagct
     2581 taggccggcc gcggccgcac ccagctttct tgtacaaagt tggcattata agaaagcatt
     2641 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgct
     2701 agggataaca gggtaattgc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2761 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2821 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2881 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2941 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3001 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3061 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3121 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3181 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3241 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3301 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3361 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3421 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3481 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3541 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     3601 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     3661 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     3721 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3781 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3841 cgcg
//
