LOCUS       CN2864_rAAV-eHGT_725m-minBglobin-SYFP2-WPRE3-BGHpA        4527 bp    DNA     circular UNA 31-AUG-2020
DEFINITION  Gibson/Homology Cloning ligation product.
ACCESSION   urn.local...4pw-ccp9eqm
VERSION     urn.local...4pw-ccp9eqm
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     misc_feature    1..141
                     /vntifkey=21
                     /locus_tag="L-ITR"
                     /ApEinfo_label="L-ITR"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="L-ITR"
     misc_feature    complement(137..469)
                     /Source="referenceToDocument: urn:local:.:4aw-ccp8h0g"
                     /label="m725-RMSYFP2seqR.ab1 (concatenated sequence 2)
                     (reversed)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    136^137
                     /Original_Bases="GAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGG"
                     /label="GAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGG"
                     /note="Geneious type: Editing History Deletion"
     ligation        137..154
                     /label="Ligation"
     primer_bind     137..155
                     /%GC="73.7"
                     /Hairpin_Tm="60.9"
                     /Self_Dimer_Tm="46.9"
                     /Tm="70.6"
                     /Pair_Dimer_Tm="None"
                     /annotation_group="17273279664438586eb6698b-adb7-41c8-835f
                     -1f8e9dc52e25"
                     /created_by="primer3"
                     /Sequence="TTCCTGCGGCCGCACGCGT"
                     /Mismatches=0
                     /Product_Size=333
                     /#_Off-target_Sites=0
                     /Off-target_Template="m725-RMSYFP2seqR.ab1 (reversed)"
                     /label="Gibson Assembly into CN1244 F'"
     misc_feature    156..445
                     /Color="#ffff55"
                     /created_by="refugio.martinez"
                     /label="eHGT725m"
     primer_bind     complement(446..469)
                     /%GC="58.3"
                     /Hairpin_Tm="51.4"
                     /Self_Dimer_Tm="2.7"
                     /Tm="65.8"
                     /Pair_Dimer_Tm="None"
                     /annotation_group="17273279664438586eb6698b-adb7-41c8-835f
                     -1f8e9dc52e25"
                     /created_by="primer3"
                     /Sequence="GACTTTTATGCCCAGCCCGAGCTC"
                     /Mismatches=0
                     /Product_Size=333
                     /#_Off-target_Sites=0
                     /Off-target_Template="m725-RMSYFP2seqR.ab1 (reversed)"
                     /label="Gibson Assembly into CN1244 R'"
     misc_feature    451..154
                     /Source="referenceToDocument: urn:local:.:3by-b7vj7bw"
                     /label="CN1244_pAAV Linearized (digested by SacI, MluI)
                     Fragment 3 (concatenated sequence 1)"
                     /note="Geneious type: Concatenated sequence"
     ligation        451..469
                     /label="Ligation"
     misc_feature    452..504
                     /vntifkey=21
                     /locus_tag="Beta\Globin\minimal\promoter"
                     /ApEinfo_label="Beta\Globin\minimal\promoter"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="Beta\Globin\minimal\promoter"
     misc_feature    461..466
                     /vntifkey=21
                     /locus_tag="TATA\box"
                     /ApEinfo_label="TATA\box"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="TATA\box"
     misc_feature    469^470
                     /Original_Bases="AGGGCAGAGCCATCTATTGCTTACATTTGCTTCTGGGANCC
                     AGNTNNNNNNNNNNNNNNNNNNN"
                     /label="AGGGCAGAGCCATCTATTGCTTACATTTGCTTCTGGGANCCAGNTNNNNN
                     NNNNNNNNNNNNNN"
                     /note="Geneious type: Editing History Deletion"
     misc_feature    493
                     /vntifkey=21
                     /locus_tag="transcriptional\start"
                     /ApEinfo_label="transcriptional\start"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="transcriptional\start"
     misc_feature    545..1264
                     /vntifkey=21
                     /locus_tag="SYFP2"
                     /ApEinfo_label="SYFP2"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="SYFP2"
     misc_feature    1299..1544
                     /vntifkey=21
                     /locus_tag="WPRE3"
                     /ApEinfo_label="WPRE3"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="WPRE3"
     misc_feature    1558..1761
                     /vntifkey=21
                     /locus_tag="BGH-pA"
                     /ApEinfo_label="BGH-pA"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="BGH-pA"
     primer          complement(1672..1691)
                     /vntifkey=27
                     /locus_tag="BGH-R"
                     /ApEinfo_label="BGH-R"
                     /ApEinfo_fwdcolor="#00ffff"
                     /ApEinfo_revcolor="#80ff00"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="BGH-R"
     misc_feature    1790..1930
                     /vntifkey=21
                     /locus_tag="R-ITR"
                     /ApEinfo_label="R-ITR"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="R-ITR"
     rep_origin      2022..2328
                     /vntifkey=33
                     /locus_tag="f1\Origin"
                     /ApEinfo_label="f1\Origin"
                     /ApEinfo_fwdcolor="#999999"
                     /ApEinfo_revcolor="#999999"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="f1\Origin"
     misc_feature    2847..3704
                     /vntifkey=21
                     /locus_tag="AmpR"
                     /ApEinfo_label="AmpR"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="AmpR"
     rep_origin      3855..4522
                     /vntifkey=33
                     /locus_tag="pUC\ori"
                     /ApEinfo_label="pUC\ori"
                     /ApEinfo_fwdcolor="#999999"
                     /ApEinfo_revcolor="#999999"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="pUC\ori"
ORIGIN      
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtttttg tgtgtctgtc ccccagtgcc
      181 gggctctgcc cctcctctgt ctttccctgc ctgctctgtg catttactct gaactcgagc
      241 aggcctattt ccaaacaccg tctagcgcgg ggctgtgcca gctccggctc tgtcaggacc
      301 aacttcccaa agtgttcgca gattctcaga gcttagctct tctcagagtt tgccattttt
      361 ttttaaaaag agatttattt tttattttaa atgatagacg ctcgtgggtg tctgtgtggg
      421 ggattgtaca catgtgtggg ggtgtgagct cgggctgggc ataaaagtca gggcagagcc
      481 atctattgct tacatttgct tctgggatcc agatctttcg aagctagcgc taccggtcgc
      541 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      601 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      661 ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc
      721 caccctcgtg accaccctgg gctacggcgt gcagtgcttc gcccgctacc ccgaccacat
      781 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      841 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
      901 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
      961 gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa
     1021 gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct
     1081 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1141 ccactacctg agctaccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     1201 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1261 gtaagtcgac ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc tggattacaa
     1321 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     1381 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     1441 cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg
     1501 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggctcgag agatcttcga
     1561 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     1621 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     1681 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     1741 gggaagacaa tagcaggcat gagatctcac gtgcggaccg agcggccgca ggaaccccta
     1801 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     1861 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     1921 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     1981 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2041 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2101 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2161 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2221 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2281 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2341 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2401 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     2461 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     2521 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     2581 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     2641 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     2701 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2761 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2821 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2881 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2941 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3001 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3061 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3121 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3181 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3241 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3301 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3361 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     3421 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3481 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3541 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3601 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3661 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3721 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3781 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3841 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3901 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3961 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4021 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4081 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4141 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4201 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4261 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4321 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4381 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4441 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4501 ccttttgctg gccttttgct cacatgt
//
