LOCUS       CN4981-pAAV-TelN-LITR-eHGT_1391h-minBG-SYFP2-P2A-3XFLAG-10aa-H2B*-WPRE3-BGHpA-RITR-TelN        5363 bp    DNA     circular UNA 25-APR-2023
DEFINITION  Generated by Gibson/Homology Cloning (Mlul/Sacl cut CN4031,
            1391h-SYFP2_5_Seq_R.ab1 (reversed)).
ACCESSION   urn.local...6j-g7vli68
VERSION     urn.local...6j-g7vli68
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     misc_feature    1..130
                     /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"
                     /vntifkey=21
                     /label="L-ITR"
     misc_feature    complement(126..646)
                     /label="1391h-SYFP2_5_Seq_R.ab1 (concatenated sequence 2)
                     (reversed)"
                     /note="Geneious type: Concatenated sequence"
     primer_bind     126..144
                     /%GC="73.7"
                     /Hairpin_Tm="60.9"
                     /Self_Dimer_Tm="46.9"
                     /Tm="70.6"
                     /annotation_group="1811417104644600e2edddbb-a1b8-4b41-9131
                     -436673e696f1"
                     /created_by="primer3"
                     /Sequence="TTCCTGCGGCCGCACGCGT"
                     /Mismatches=0
                     /#_Local_Off-target_Sites=0
                     /Local_Off-target="1391h-SYFP2_5_Seq_R.ab1"
                     /label="CN1244 F"
     misc_feature    125^126
                     /Original_Bases="AGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGG
                     GG"
                     /label="AGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGG"
                     /note="Geneious type: Editing History Deletion"
     ligation        126..143
                     /label="Ligation"
     regulatory      145..622
                     /created_by="erin.groce"
                     /label="eHGT_1391h"
     primer_bind     complement(623..646)
                     /%GC="58.3"
                     /Hairpin_Tm="51.4"
                     /Self_Dimer_Tm="2.7"
                     /Tm="65.8"
                     /annotation_group="1811417032588000fed2fb7a-e147-48cf-bb50
                     -a9bc6f381fa9"
                     /created_by="primer3"
                     /Sequence="GACTTTTATGCCCAGCCCGAGCTC"
                     /Mismatches=0
                     /#_Local_Off-target_Sites=0
                     /Local_Off-target="1391h-SYFP2_5_Seq_R.ab1"
                     /label="CN 1244R"
     ligation        628..646
                     /label="Ligation"
     misc_feature    628..143
                     /label="Mlul/Sacl cut CN4031 (concatenated sequence 1)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    629..681
                     /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"
                     /vntifkey=21
                     /label="Beta\Globin\minimal\promoter"
     misc_feature    638..643
                     /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"
                     /vntifkey=21
                     /label="TATA\box"
     misc_feature    646^647
                     /Original_Bases="AGGGCAGAGCCATCTATTGCTTACATTTGCTTCTGGGANCC
                     AGNNCNNNNNNNNNNNNNNNNNNNNNNN"
                     /label="AGGGCAGAGCCATCTATTGCTTACATTTGCTTCTGGGANCCAGNNCNNNN
                     NNNNNNNNNNNNNNNNNNN"
                     /note="Geneious type: Editing History Deletion"
     misc_feature    670
                     /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"
                     /vntifkey=21
                     /label="transcriptional\start"
     misc_feature    722..1436
                     /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"
                     /vntifkey=21
                     /label="SYFP2"
     misc_feature    1439..1516
                     /locus_tag="P2A"
                     /ApEinfo_label="P2A"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /vntifkey=21
                     /label="P2A"
     CDS             1517..1582
                     /codon_start=1
                     /product="three tandem FLAG epitope tags, followed by an
                     enterokinase cleavage site"
                     /locus_tag="3xFLAG"
                     /ApEinfo_label="3xFLAG"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /vntifkey=4
                     /label="3xFLAG"
     misc_feature    1583..1612
                     /locus_tag="10\a.a.\linker"
                     /ApEinfo_label="10\a.a.\linker"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /vntifkey=21
                     /label="10\a.a.\linker"
     misc_feature    1613..1987
                     /locus_tag="H2B"
                     /ApEinfo_label="H2B"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /vntifkey=21
                     /label="H2B"
     misc_feature    2022..2267
                     /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"
                     /vntifkey=21
                     /label="WPRE3"
     misc_feature    2022..2181
                     /locus_tag="gamma"
                     /ApEinfo_label="gamma"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /vntifkey=21
                     /label="gamma"
     misc_feature    2187..2267
                     /locus_tag="alpha"
                     /ApEinfo_label="alpha"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /vntifkey=21
                     /label="alpha"
     misc_feature    2281..2484
                     /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"
                     /vntifkey=21
                     /label="BGH-pA"
     primer          complement(2395..2414)
                     /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"
                     /vntifkey=27
                     /label="BGH-R"
     misc_feature    2513..2653
                     /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"
                     /vntifkey=21
                     /label="R-ITR"
     misc_feature    2660..2715
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     unsure          2660..2715
                     /created_by="refugio.martinez"
                     /Color="#ffddff"
                     /modified_by="refugio.martinez"
                     /label="TelN_seq"
     rep_origin      2801..3107
                     /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"
                     /vntifkey=33
                     /label="f1\Origin"
     misc_feature    3626..4483
                     /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"
                     /vntifkey=21
                     /label="AmpR"
     rep_origin      4634..5301
                     /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"
                     /vntifkey=33
                     /label="pUC\ori"
     misc_feature    5302..5358
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     unsure          5302..5357
                     /created_by="refugio.martinez"
                     /Color="#ffddff"
                     /modified_by="refugio.martinez"
                     /label="TelN_seq"
ORIGIN      
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcactgt gatgcttcta agagctttaa acttgtaaat
      181 aacccctccc agaagcgcaa atctccatat cttaatatca tgaaagctgg aactgtaaaa
      241 taaaatctgt tagcagctga aatcttaatg aggttctaca aagaagaggc ttttctgcaa
      301 tgactcaatg catggaaaca gttgagtaat tataaatgca tgggctgaat gtgagttggc
      361 aaatataata ccacagaatg tacccagaac tagggttgtc ccagttcaaa tgagactata
      421 ttcctttacc agtacaggca aacaactaac ctgaaatgtc tttggctaag agataggtgt
      481 gagcccataa aattatgtcc cttgactcct catttacatg gtgacaatct gttctggccc
      541 accagcagcc tgttcaacaa caaagcactt cagaaagagc tctgcataat attggtttca
      601 tcagacataa tagaagccaa tggagctcgg gctgggcata aaagtcaggg cagagccatc
      661 tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac cggtcgccac
      721 catggtctcc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      781 cggcgatgtc aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
      841 cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac
      901 cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa
      961 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     1021 cttcaaagac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1081 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1141 caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa
     1201 cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc
     1261 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1321 ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     1381 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg
     1441 ctctggtgct accaacttct cactgttgaa acaggcaggg gatgtagagg agaatccagg
     1501 gcctggtgct agtggagact acaaagacca tgacggagat tataaagatc atgacatcga
     1561 ttacaaggat gacgatgaca agtccggact cagatctgga ggctccggag gcccagagcc
     1621 agcgaagtct gctcccgccc cgaaaaaggg ctccaagaag gcggtgacta aggcgcagaa
     1681 gaaaggcggc aagaagcgca agcgcagccg caaggagagc tattccatct atgtgtataa
     1741 ggttctgaag caggtccacc ctgacaccgg catttcgtcc aaggccatgg gcatcatgaa
     1801 ctcgtttgtg aacgacattt tcgagcgcat cgcaggtgag gcttcccgcc tggcgcatta
     1861 caacaagcgc tcgaccatca cctccaggga gatccagacg gccgtgcgcc tgctgctgcc
     1921 tggggagttg gccaagcacg ccgtgtccga gggtactaag gccatcacca agtacaccag
     1981 cgctaagtaa tgaggcgcgc cgcggccgcg aattcgatat cataatcaac ctctggatta
     2041 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2101 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2161 ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc
     2221 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc gagagatctt
     2281 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2341 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2401 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2461 attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc gcaggaaccc
     2521 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     2581 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     2641 agctgcctgc aggggcgcct atcagcacac aattgcccat tatacgcgcg tataatggac
     2701 tattgtgtgc tgatatgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2761 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2821 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2881 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2941 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     3001 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3061 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3121 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3181 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     3241 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3301 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3361 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3421 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3481 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3541 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3601 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3661 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3721 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3781 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3841 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3901 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3961 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4021 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4081 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4141 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4201 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4261 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4321 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4381 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4441 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4501 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4561 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4621 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4681 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4741 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4801 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4861 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4921 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4981 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5041 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5101 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5161 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5221 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5281 cttttgctgg ccttttgctc atatcagcac acaattgccc attatacgcg cgtataatgg
     5341 actattgtgt gctgataaca tgt
//
