LOCUS       Exported                5966 bp ds-DNA     circular UNA 27-APR-2020
DEFINITION  Golden Gate ligation product.
ACCESSION   urn.local...ci-buo3vji
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5966)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, May 13, 2020 from SnapGene Viewer 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5966
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     misc_feature    1..5320
                     /label=pAGM22082_cRED  extraction (concatenated sequen
                     /label=pAGM22082_cRED  extraction (concatenated sequence
                     1)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:2n-9725dlj"
     misc_feature    1..4462
                     /label=Ligated sequences 3 extraction (concatenated
                     se
                     /label=Ligated sequences 3 extraction (concatenated
                     sequence 1)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:b2-9489ukd"
     misc_feature    1..4339
                     /label=pAGM22082 extraction (concatenated sequence 1)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:cl-buo3vjr"
     terminator      15..143
                     /label=T7 terminator
     misc_feature    complement(57..75)
                     /label=T7 Terminal primer
     primer_bind     complement(75..99)
                     /label=Sequencing primer
     misc_feature    155..198
                     /label=beginining of bacteriophage seq
     rep_origin      172..478
                     /label=f1 origin
     CDS             complement(703..1518)
                     /label=KanR2 CDS
     misc_feature    1075
                     /label=A>T esprem
     rep_origin      1624..2243
                     /label=pBR322 origin
     misc_feature    2639
                     /label=T>A esprem
     misc_feature    2642..2664
                     /label=pGEX 3 primer
     misc_feature    complement(2658..2882)
                     /label=rop large?
     misc_feature    complement(2658..2849)
                     /label=ROP
     CDS             2881..3246
                     /label=mob?
     misc_feature    3159
                     /label=C>G bpirem
     misc_feature    complement(3249..3602)
                     /label=hyp prot
     misc_feature    3522
                     /label=G>C bpirem
     CDS             complement(3658..4339)
                     /label=ORF frame 1
     misc_feature    complement(3658..4339)
                     /label=lacI
     misc_feature    3778
                     /label=G>C esprem
     misc_feature    3910
                     /label=T>C bpirem
     misc_feature    4249
                     /label=T>C bpirem
     CDS             complement(4336..4462)
                     /label=ORF frame 1
     misc_feature    4336..4462
                     /label=Digestion of pAGM22082_Dummy by BstXI - Fragmen
                     /label=Digestion of pAGM22082_Dummy by BstXI - Fragment B 
                     (concatenated sequence 2)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:ap-9489tcx"
     misc_feature    4336..4462
                     /label=pAGM22082 extraction (concatenated sequence 1)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:co-buo3vjr"
     misc_feature    complement(4336..4462)
                     /label=lacI
     misc_feature    4336..4339
                     /label=Ligation
                     /label=nonstandard type:  ligation
     CDS             complement(4459..4591)
                     /label=ORF frame 1
     misc_feature    4459..4591
                     /label=Digestion of pAGM22082_Dummy by BstXI - Fragmen
                     /label=Digestion of pAGM22082_Dummy by BstXI - Fragment D 
                     (concatenated sequence 2)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:b8-9489vuz"
     misc_feature    4459..4591
                     /label=pAGM22082 extraction (concatenated sequence 1)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:cn-buo3vjr"
     misc_feature    complement(4459..4591)
                     /label=lacI
     misc_feature    4459..4462
                     /label=Ligation
                     /label=nonstandard type:  ligation
     misc_feature    4554..4591
                     /label=end of lacI homologous to e.coli genome
     misc_feature    4588..5320
                     /label=Ligated sequences 2 (concatenated sequence 1)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:aj-9489q9n"
     misc_feature    4588..5320
                     /label=Ligated sequences 3 - Ligated sequences 2
                     (conc
                     /label=Ligated sequences 3 - Ligated sequences 2
                     (concatenated sequence 1) (concatenated sequence 3)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:b2-9489ukd"
     misc_feature    4588..5320
                     /label=Digestion of pAGM22082_Dummy by BstXI - Fragmen
                     /label=Digestion of pAGM22082_Dummy by BstXI - Fragment C 
                     (concatenated sequence 1)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:a4-9489p6m"
     misc_feature    4588..5320
                     /label=pAGM22082 extraction (concatenated sequence 1)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:cm-buo3vjr"
     misc_feature    4588..4826
                     /label=end of lacI homologous to e.coli genome
     misc_feature    complement(4588..4749)
                     /label=lacI
     CDS             complement(4588..4617)
                     /label=ORF frame 1
     misc_feature    4588..4591
                     /label=Ligation
                     /label=nonstandard type:  ligation
     misc_feature    5024..5043
                     /label=pBRrevBam primer
     misc_feature    5127..5145
                     /label=T7 promoter
                     /note="Geneious type: promoter prokaryotic"
     misc_feature    5145..5172
                     /label=lacO
     misc_feature    5192..5208
                     /label=T7 transl en RBS
     misc_feature    5202..5226
                     /label=Sequencing primer
                     
                     /note="Geneious type: primer_bind_reverse"
                     /note="/%GC=52.0"
                     /note="/Hairpin_Tm=None"
                     /note="/Self_Dimer_Tm=18.1"
                     /note="/Sequence=GAAGGAGATATACCATGGGCAGCAG"
                     /note="/Tm=62.8"
                     /note="/created_by=primer3"
     misc_feature    5228..5245
                     /label=N-ter 6xHis
     misc_feature    5275..5293
                     /label=Xpress fwd primer
     misc_feature    5276..5308
                     /label=T7 leader tag
     misc_feature    5317..5966
                     /label=sfGFP1-10opt (concatenated sequence 2)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:bs-buo3rqb"
     misc_feature    5317..5320
                     /label=Ligation
                     /label=nonstandard type:  ligation
     misc_feature    5317..5320
                     /label=Geneious type: Editing History Insertion
                     /note="Geneious type: Editing History Insertion"
                     /note="/Original_Bases="
     misc_feature    5317..5320
                     /label=Digestion of Dummy pAGM cloning by BsaI - Fragm
                     /label=Digestion of Dummy pAGM cloning by BsaI - Fragment
                     2 (concatenated sequence 2)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:cp-buo3vjr"
     misc_feature    join(5963..5966,1..4339)
                     /label=Ligated sequences 2 (concatenated sequence 1)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:aj-9489q9n"
     misc_feature    join(5963..5966,1..4339)
                     /label=Ligated sequences (concatenated sequence 2)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:9y-9489m57"
     misc_feature    join(5963..5966,1..4339)
                     /label=Digestion of pAGM22082_Dummy by BstXI -Fragment
                     /label=Digestion of pAGM22082_Dummy by BstXI -Fragment A 
                     (concatenated sequence 2)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:6k-9488c8y"
     misc_feature    5963..5966
                     /label=Ligated sequences 3 - Ligated sequences 2
                     (conc
                     /label=Ligated sequences 3 - Ligated sequences 2
                     (concatenated sequence 1) (concatenated sequence 3)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:b2-9489ukd"
     misc_feature    5963..5966
                     /label=Ligation
                     /label=nonstandard type:  ligation
     misc_feature    5963..5966
                     /label=pAGM22082_cRED  - BsaI overhang (concatenated
                     s
                     /label=pAGM22082_cRED  - BsaI overhang (concatenated
                     sequence 3)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:2n-9725dlj"
     misc_feature    5963..5966
                     /label=Geneious type: Editing History Insertion
                     /note="Geneious type: Editing History Insertion"
                     /note="/Original_Bases="
     misc_feature    5963..5966
                     /label=Digestion of Dummy pAGM cloning by BsaI - Fragm
                     /label=Digestion of Dummy pAGM cloning by BsaI - Fragment
                     2 (concatenated sequence 2)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:cq-buo3vjr"
     misc_feature    5963..5966
                     /label=pAGM22082 - BsaI overhang (concatenated sequenc
                     /label=pAGM22082 - BsaI overhang (concatenated sequence
                     3)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:cl-buo3vjr"
ORIGIN
        1 gggatgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
       61 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
      121 gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc
      181 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
      241 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
      301 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
      361 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
      421 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
      481 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
      541 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
      601 tttacaattt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
      661 ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag
      721 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
      781 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
      841 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
      901 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
      961 caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     1021 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagtcgaaa
     1081 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     1141 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     1201 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     1261 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     1321 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     1381 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     1441 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc
     1501 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     1561 tgttcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1621 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1681 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1741 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1801 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1861 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1921 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1981 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2041 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     2101 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     2161 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     2221 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     2281 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     2341 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     2401 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2461 gcatatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     2521 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     2581 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgac
     2641 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     2701 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     2761 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     2821 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
     2881 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     2941 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     3001 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
     3061 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
     3121 gacttccgcg tttccagact ttacgaaaca cggaaaccca agaccattca tgttgttgct
     3181 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
     3241 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
     3301 atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     3361 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     3421 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     3481 agcgctgccg gcacctgtcc tacgagttgc atgataaaga acacagtcat aagtgcggcg
     3541 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     3601 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     3661 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3721 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgacac
     3781 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     3841 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     3901 tgagctgtcc tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
     3961 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     4021 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     4081 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     4141 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     4201 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtcct catgggagaa
     4261 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     4321 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     4381 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     4441 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     4501 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     4561 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     4621 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     4681 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     4741 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     4801 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     4861 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
     4921 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     4981 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     5041 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     5101 ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
     5161 taacaattcc cctctagaaa taattttgcc tttaacttta agaaggagat ataccatggg
     5221 cagcagccat catcatcatc atcacagcag cggcctggtg ccgcgcggca gccatatggc
     5281 tagcatgact ggtggacagc aaatgggtcg ggatggaatg agtaaaggcg aagagctgtt
     5341 cactggcgtt gttccgattc tggtcgaact ggatggtgat gttaacggcc acaaattcag
     5401 cgtccgtggg gaaggcgagg gtgatgctac aattggcaaa ctgaccctga aattcatctg
     5461 taccaccggg aaactgcctg ttccttggcc tacactggtt accactctga cctatggcgt
     5521 gcaatgtttt agccgctatc ctgatcacat gaaacgtcac gatttcttca aaagcgctat
     5581 gcctgaaggt tatgtccaag agcgtaccat cagctttaaa gacgatggca aatataaaac
     5641 ccgtgccgtg gtgaaattcg aaggtgatac cctggtcaac cgtattgaac tgaaagggac
     5701 cgatttcaaa gaggacggca acattctggg ccataaactg gagtataact tcaacagcca
     5761 taacgtgtac attaccgccg acaaacagaa aaacgggatc aaagcgaact tcacagtccg
     5821 ccacaacgtg gaagatggtt ccgttcagct ggcagatcat tatcagcaga acaccccgat
     5881 tggagatggt cctgtactgc tgccggacaa tcactatctg agcacccaga cagtgctgag
     5941 taaagacccg aacgaaaaat aagctt
//
