LOCUS       40924_6347        7024 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Transformation vector pBHRF-108, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7024)
  AUTHORS   Moody LA, Kelly S, Coudert Y, Nimchuk ZL, Harrison CJ, Langdale JA.
  TITLE     Somatic hybridization provides segregating populations for the 
            identification of causative mutations in sterile mutants of the moss
            Physcomitrella patens
  JOURNAL   New Phytol. (2018) In press
  PUBMED    29498048
REFERENCE   2  (bases 1 to 7024)
  AUTHORS   Schaefer DG.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-JAN-2018) Institute of Biology, University of 
            Neuchatal, Rue Emile-Argand 11, Neuchatal CH-2000, Switzerland
REFERENCE   3  (bases 1 to 7024)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7024)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "New Phytol.
            (2018) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-JAN-2018) Institute of Biology, University of Neuchatal, Rue 
            Emile-Argand 11, Neuchatal CH-2000, Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7024
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     142..158
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    182..1293
                     /note="108 targeting locus for ectopic integration of 
                     transgenes"
     protein_bind    1363..1396
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1599..1944
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             1977..3002
                     /label=HygR
                     /note="hygromycin B phosphotransferase"
     regulatory      3018..3198
                     /label=35S terminator
                     /note="35S terminator"
                     /regulatory_class="terminator"
     protein_bind    complement(3276..3309)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3366..4364
                     /note="108 targeting locus for ectopic integration of 
                     transgenes"
     promoter        complement(4383..4401)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4416..4432)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4440..4456)
                     /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)."
     promoter        complement(4464..4494)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4509..4530)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4818..5406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5580..6437)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6438..6542)
                     /label=AmpR promoter
     rep_origin      6569..7024
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
      121 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctaatt accctgttat
      181 ccctaggagt gtgcaaccaa ataccgaccc tgcacctatc gaagacacag cgatcatatc
      241 aaatcacata tttgacatgt caagatgcaa acttgttatc tacacaaaat tgacctaaga
      301 tataattaag cctatagagc tccatatcat aggcttactc atgaaaacat caactcacgg
      361 actgtcgtct ttcaagaaga acgttagcat gaatgttcca caataaacaa aaagaaaagt
      421 ttagaagtaa ggacagatac catcgacact ggagaaaata gccatccgta gacgatgcct
      481 cggtaacatc ttcaggagaa agactcgaat ctccccgtcg caacagcaca taggccgaca
      541 actcgcagcc tacaatggga atgtcggtcg ggaggtgcac ccgcactaca ctcagcattc
      601 ttccccgctt tcgccccctc ccttatctcc agctccgctt aagatgatac tcacgctttt
      661 gtacttcggc tctttcactt ccagacgaat ttcatgaaaa gttacatgtg cttgaggccg
      721 atcagccatc aggcgccccc tgtgcgcgcc agcgttctcc cctcaacctc gccccctctc
      781 actctttgaa aagtgtgtgt ctaaatctgg cagtggcgat ccctcgaaac ctaaaactct
      841 tgcgaaacat aagtaactta agttttcctc tacaaaactt gcaagtacga catgcaccca
      901 cacaagtgca acacggacgc ctcttgtttt ctctacattc tcatagtaga acatgagatt
      961 cctctcagaa acgcattccc tcccattcac ttaatcccct gttccacgcg tgtcagggag
     1021 tactcttgca atcataaagc gatggcttgg tcattcaaca cttggtagct tcctgcaaat
     1081 aaaccctaaa gttgtcagag cacttgagga gaagctcttc cctcactcca cctattcaca
     1141 ctacctttcc acttgagaag gacagctgct gcagcaaacc cccaaaaaaa aatagggagc
     1201 gggaggggaa caataaatgc attacctaaa ccacgcaacc atctctcatc gaacaagcct
     1261 caccgcaacg cagactcttc ctccaacacc cctgtcgact ctagaggatc ccacgtgccc
     1321 gggcgtttaa accaattgtt cgaacgtacg tcgcgactcg agataacttc gtataatgta
     1381 tgctatacga agttatggta ccgagctcga attcccatgg agtcaaagat tcaaatagag
     1441 gacctaacag aactcgccgt aaagactggc gaacagttca tacagagtct cttacgactc
     1501 aatgacaaga agaaaatctt cgtcaacatg gtggagcacg acacgcttgt ctactccaaa
     1561 aatatcaaag atacagtctc agaagaccaa agggcaattg agacttttca acaaagggta
     1621 atatccggaa acctcctcgg attccattgc ccagctatct gtcactttat tgtgaagata
     1681 gtggaaaagg aaggtggctc ctacaaatgc catcattgcg ataaaggaaa ggccatcgtt
     1741 gaagatgcct ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg
     1801 gaaaaagaag acgttccaac cacgtcttca aagcaagtgg attgatgtga tatctccact
     1861 gacgtaaggg atgacgcaca atcccactat ccttcgcaag acccttcctc tatataagga
     1921 agttcatttc atttggagag gacagggtac ccggggatcc ccgggggcaa tgagatatga
     1981 aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg
     2041 tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag
     2101 gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt
     2161 atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg
     2221 aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag
     2281 acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga
     2341 tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg
     2401 gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact
     2461 ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga
     2521 tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca
     2581 acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt
     2641 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta
     2701 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc
     2761 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca
     2821 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg
     2881 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg
     2941 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccggga tcctctagag
     3001 tcgacctgca ggcatgccgc tgaaatcacc agtctctctc tacaaatcta tctctctcta
     3061 taataatgtg tgagtagttc ccagataagg gaattagggt tcttataggg tttcgctcat
     3121 gtgttgagca tataagaaac ccttagtatg tatttgtatt tgtaaaatac ttctatcaat
     3181 aaaatttcta attcctaaaa ccaaaatcca gtgggtaccg agctcgaatt cgcggccgcg
     3241 ggcccatatg gcccaccggt gggccggcca gatctataac ttcgtataat gtatgctata
     3301 cgaagttata ctagtggcgc cacgcgtgat atcatgcatg ttaacatcga tccatgggcg
     3361 cgcctcgtac cacaccactt ccaggctcag aagcagaatc gattcagcag tcactaaccc
     3421 tctctatgcg acagccacga gaattacaaa ctgggtctgc cggacttaca tatcaatcaa
     3481 cggaatcacg tccaagtaaa ataatccagc gaacttatcc acataggatc tgagggggga
     3541 acgtgaatga gccaggtcta cacgcagcat gggagagaag gggggggggg gttttcaaat
     3601 agagtccaaa attagcccaa acgagggagg gagaatgaat gaggctgtcg agggcgggcg
     3661 gagatggaac gaagagtcgg gggggggggg ggatagcgaa ggcgaagagg ctctcaccca
     3721 cccattgcgc ttctttgcga actcgaccac cgaatctctc cctcccttcc ctcttcctct
     3781 ccctctcctc tctctctctc tctcgtatgt gtgtgtgtgt ctaaattcgc ccaattcgtc
     3841 gtcccttccg ccatcgccat tgccatgcca atcaatgcct tagaaacggg cactaccatg
     3901 gcatcccccc ctttgcatcc ttggaccctc cggagggagg gagggagtca acgttcacac
     3961 tccgaagtca acctggtcgc tagacttttg gcttcctctg tttatctttt gtctcggtag
     4021 atccctcttg gaatatatgg atggatggat gaaattatgg atgaatgatt gaatggcatt
     4081 tgtgaaccat tcgatccctc ctgagtgcct gtgtccgtca gtccgtgtaa ctgtgactga
     4141 ctgttctatc tcatcatcca tcctttcttc atgctctact cggactgaac tctctctctc
     4201 tctctctctc tctctctcgc tctgtgtatg tgtgtgtgtg tttgtttatt gacttctgaa
     4261 cttgtcattt ggttctaatg tagagtgaaa gtcgtttaaa gcagaatatg caaatcgtgc
     4321 attttaagtg gtgtaatgta atgtattttg gggtggtgat gtcattaatt aaatttaaat
     4381 tccctatagt gagtcgtatt aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4441 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4501 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4561 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4621 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4681 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4741 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4801 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4861 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4921 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4981 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5041 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5101 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5161 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5221 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     5281 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5341 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5401 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5461 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5521 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5581 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5641 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5701 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5761 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5821 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5881 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5941 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6001 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6061 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6121 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6181 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6241 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6301 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6361 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6421 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6481 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6541 cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg
     6601 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     6661 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     6721 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     6781 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     6841 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     6901 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     6961 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     7021 attt
//
