LOCUS       CN5028-pAAV-TelN-LITR-eHGT_1404m-minBG-SYFP2-P2A-3XFLAG-10aa-H2B*-WPRE3-BGHpA-RITR-TelN        5752 bp    DNA     circular UNA 16-JUN-2023
DEFINITION  Generated by Gibson/Homology Cloning (Mlul/Sacl cut CN4031,
            1404.2-SYFP2_5_seq_R.ab1 (reversed)).
ACCESSION   urn.local...g5-gh1w8al
VERSION     urn.local...g5-gh1w8al
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..1035)
                     /label="1404.2-SYFP2_5_seq_R.ab1 (concatenated sequence 2)
                     (reversed)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    126..130
                     /Original_Bases="NANNNNNNNNNNNNNNNNNAGGGAGTGGCCAACTCCATCAN
                     TAGGGGNNNN"
                     /label="NANNNNNNNNNNNNNNNNNAGGGAGTGGCCAACTCCATCANTAGGGGNNN
                     N"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     126..144
                     /%GC="73.7"
                     /Hairpin_Tm="60.9"
                     /Self_Dimer_Tm="46.9"
                     /Tm="70.6"
                     /annotation_group="6128216113622002f786b78-8c75-427e-b6ba-
                     d40d905f29b9"
                     /created_by="primer3"
                     /Sequence="TTCCTGCGGCCGCACGCGT"
                     /Mismatches=0
                     /#_Local_Off-target_Sites=0
                     /Local_Off-target="1404.2-SYFP2_5_seq_R.ab1 (reversed)"
                     /label="CN1244 F"
     ligation        126..143
                     /label="Ligation"
     regulatory      145..1011
                     /created_by="erin.groce"
                     /label="eHGT_1404m"
     misc_feature    174..177
                     /Original_Bases="NNN"
                     /label="NNN"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    198^199
                     /Original_Bases="T"
                     /label="T"
                     /note="Geneious type: Editing History Deletion"
     misc_feature    316
                     /Original_Bases="N"
                     /label="N"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     complement(1012..1035)
                     /%GC="58.3"
                     /Hairpin_Tm="51.4"
                     /Self_Dimer_Tm="2.7"
                     /Tm="65.8"
                     /annotation_group="612760451317900572028bb-1fa8-40f9-b1b3-
                     43c113853cbc"
                     /created_by="primer3"
                     /Sequence="GACTTTTATGCCCAGCCCGAGCTC"
                     /Mismatches=0
                     /#_Local_Off-target_Sites=0
                     /Local_Off-target="1404.2-SYFP2_5_seq_R.ab1"
                     /label="CN 1244R"
     ligation        1017..1035
                     /label="Ligation"
     misc_feature    1017..143
                     /label="Mlul/Sacl cut CN4031 (concatenated sequence 1)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    1018..1070
                     /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    1027..1032
                     /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    1035^1036
                     /Original_Bases="AGGGCAGAGCCATCTATTGCTTACATTTGCTTCTGGGANCC
                     AGNNNNNNGANNNNNNNNNNNNNNNNN"
                     /label="AGGGCAGAGCCATCTATTGCTTACATTTGCTTCTGGGANCCAGNNNNNNG
                     ANNNNNNNNNNNNNNNNN"
                     /note="Geneious type: Editing History Deletion"
     misc_feature    1059
                     /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    1111..1825
                     /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    1828..1905
                     /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             1906..1971
                     /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    1972..2001
                     /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    2002..2376
                     /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    2411..2656
                     /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    2411..2570
                     /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    2576..2656
                     /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    2670..2873
                     /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(2784..2803)
                     /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    2902..3042
                     /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    3049..3104
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     unsure          3049..3104
                     /created_by="refugio.martinez"
                     /Color="#ffddff"
                     /modified_by="refugio.martinez"
                     /label="TelN_seq"
     rep_origin      3190..3496
                     /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    4015..4872
                     /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      5023..5690
                     /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    5691..5747
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     unsure          5691..5746
                     /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 gcgtatgtat aagatgacta ctggcctata tttacctgac
      181 aagaatctat tcctccttct tcatatcctg cacatatcat actttcagtg atgttgtatt
      241 ctggcagctg ttgctggcac ttttcattgg atataagagg aacatcagct tctttcagca
      301 catcaacagt agaacctgct tgaaatggga aatgcagaca gccagtcagt aatgattaac
      361 aggcatcctc gaatgaaacg ccatccatct gatttatccc agcattttgc tctcattatc
      421 aagtactatg tggatgtata atggaactaa ctaccaacct tgcactttca agtacataat
      481 aaacccatct gtgcactttg gtaaatggag agcatttctt tcttcctttc ctaaggggta
      541 gtaagaattg tggattggac tagggtggtc ttttaggcat tctcagtatt ttttaaagag
      601 ttacagtgtt gctggtagaa aggaagtctg ccagcacaaa gtgttcattc atggaaatca
      661 gcacagattt ggatatccag aagagctttt catccagcac ctgaggaggt gctccctcct
      721 gtataaaggc attcagctcc tttgtggttc tgtgtgtgcg agagcaggtg cacgctgaac
      781 aggggacaaa gccagctgaa ctgtgctcta ctcttggggc aatatgcttg ccatcccagg
      841 caggtgaaga gaaaggaagc atatttcagc attgtgtgtt tgcatttgtt ttatcacaaa
      901 tcactcactg caaggatgaa cctctcctta ctaaactact gttcccaaga agcttgtgaa
      961 gcaagcaggt gttagtacct tcagaaaagg aacataagca tttattcatg agagctcggg
     1021 ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat
     1081 ctttcgaagc tagcgctacc ggtcgccacc atggtctcca agggcgagga gctgttcacc
     1141 ggggtggtgc ccatcctggt cgagctggac ggcgatgtca acggccacaa gttcagcgtg
     1201 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc
     1261 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag
     1321 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1381 gaaggctacg tccaggagcg caccatcttc ttcaaagacg acggcaacta caagacccgc
     1441 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1501 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1561 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     1621 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1681 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa
     1741 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1801 actctcggca tggacgagct gtacaagggc tctggtgcta ccaacttctc actgttgaaa
     1861 caggcagggg atgtagagga gaatccaggg cctggtgcta gtggagacta caaagaccat
     1921 gacggagatt ataaagatca tgacatcgat tacaaggatg acgatgacaa gtccggactc
     1981 agatctggag gctccggagg cccagagcca gcgaagtctg ctcccgcccc gaaaaagggc
     2041 tccaagaagg cggtgactaa ggcgcagaag aaaggcggca agaagcgcaa gcgcagccgc
     2101 aaggagagct attccatcta tgtgtataag gttctgaagc aggtccaccc tgacaccggc
     2161 atttcgtcca aggccatggg catcatgaac tcgtttgtga acgacatttt cgagcgcatc
     2221 gcaggtgagg cttcccgcct ggcgcattac aacaagcgct cgaccatcac ctccagggag
     2281 atccagacgg ccgtgcgcct gctgctgcct ggggagttgg ccaagcacgc cgtgtccgag
     2341 ggtactaagg ccatcaccaa gtacaccagc gctaagtaat gaggcgcgcc gcggccgcga
     2401 attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2461 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2521 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc
     2581 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2641 actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt
     2701 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2761 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2821 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc
     2881 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     2941 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     3001 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgccta tcagcacaca
     3061 attgcccatt atacgcgcgt ataatggact attgtgtgct gatatgatgc ggtattttct
     3121 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     3181 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3241 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3301 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3361 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     3421 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3481 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     3541 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3601 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     3661 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3721 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3781 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     3841 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3901 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3961 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4021 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4081 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4141 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4201 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4261 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4321 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4381 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4441 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4501 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4561 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4621 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4681 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4741 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4801 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4861 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4921 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4981 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5041 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5101 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5161 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     5221 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5281 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5341 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5401 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5461 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5521 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5581 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5641 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca tatcagcaca
     5701 caattgccca ttatacgcgc gtataatgga ctattgtgtg ctgataacat gt
//
