LOCUS       62056_10895        4628 bp DNA     circular SYN 01-FEB-2020
DEFINITION  Cloning vector pFLGA1, complete sequence.
ACCESSION   MN043928
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4628)
  AUTHORS   Gunadi A, Finer J.
  TITLE     CRISPRi validation of regulatory elements in a soybean (Glycine max)
            elongation factor-1 alpha promoter
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4628)
  AUTHORS   Gunadi A, Finer J.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JUN-2019) Horticulture and Crop Science, The Ohio 
            State University, 216 Williams Hall, 1680 Madison Ave., Wooster, OH 
            44691, USA
REFERENCE   3  (bases 1 to 4628)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-JUN-2019) Horticulture and Crop Science, The Ohio State 
            University, 216 Williams Hall, 1680 Madison Ave., Wooster, OH 44691,
            USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4..20)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        494..598
                     /label=AmpR promoter
     CDS             599..1456
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1630..2218
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2506..2527
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2542..2572
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2580..2596
                     /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     2604..2620
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      2695..3046
                     /label=U6.6 Promoter from Medicago truncatula
                     /note="U6.6 Promoter from Medicago truncatula"
                     /regulatory_class="promoter"
     tRNA            3053..3129
                     /product="tRNA-Gly"
                     /label=plant tRNA (Gly)
                     /note="plant tRNA (Gly)"
     CDS             3407..4195
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     misc_RNA        4350..4425
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     regulatory      4426..4617
                     /label=U6-26 pol III terminator from Arabidopsis thaliana
                     /note="U6-26 pol III terminator from Arabidopsis thaliana"
                     /regulatory_class="terminator"
ORIGIN
        1 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
       61 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
      121 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct
      181 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
      241 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
      301 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
      361 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
      421 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
      601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1141 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtggttctcg
     1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct
     2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2341 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2401 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     2461 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     2521 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     2581 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttat
     2641 taccctgtta tccctagata tcgattttgg ctaccttaac tctcttaagg tagcatgcct
     2701 atcttatatg atcaatgagg catttaattg ggtgcatatg atggtgaaaa aaggtgcagc
     2761 tcctggcttg ggaatgatga ctcatgtgga atttggtctt aaatttatca catccttttg
     2821 ggatgtgatg attgtatcac ttgttcattt tgcaaagaca aggtgcactg ctacaaactt
     2881 tggtttaatc tgaaataaaa caaaactcac tgagaggaag atgcatccca gtaggtgaaa
     2941 gtcgagaagg atttgcatgt tactattaca cttgcttttt agtcccacat cgtctgaaac
     3001 ataaaatatt tcagcgttta aatacttcaa gcgaaccagt aggcttccta ggaacaaagc
     3061 accagtggtc tagtggtaga atagtaccct gccacggtac agacccgggt tctattcccg
     3121 gctggtgcag agaccaaccc agtggacata agcctgttcg gttcgtaagc tgtaatgcaa
     3181 gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg
     3241 cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt ctatgcctcg
     3301 ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa
     3361 cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacatcatgg gggaagcggt
     3421 gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc gccatctcga
     3481 accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc tgaagccaca
     3541 cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc
     3601 tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga ttctccgcgc
     3661 tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc cagctaagcg
     3721 cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct tcgagccagc
     3781 cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata gcgttgcctt
     3841 ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc tatttgaggc
     3901 gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg atgagcgaaa
     3961 tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa tcgcgccgaa
     4021 ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc ccgtcatact
     4081 tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc gcgcagatca
     4141 gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg gcaaataatg
     4201 tctagctaga aattcgttca agccgacgcc gcttcgcggc gcggcttaac tcaagcgtta
     4261 gatgcactaa gcacataatt gctcacagcc aaactatcag gtcaagtctg cttttattat
     4321 ttttaagcgt gcataataag ccggtctcgg ttttagagct agaaatagca agttaaaata
     4381 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tttgcaaaat
     4441 tttccagatc gatttcttct tcctctgttc ttcggcgttc aatttctggg gttttctctt
     4501 cgttttctgt aactgaaacc taaaatttga cctaaaaaaa atctcaaata atatgattca
     4561 gtggttttgt acttttcagt tagttgagtt ttgcagttcc gatgagataa accaataagc
     4621 ttgcatgc
//
