LOCUS       PB-3'-FT-ENGRAM         5679 bp ds-DNA     circular     14-NOV-2021
DEFINITION  .
FEATURES             Location/Qualifiers
     primer_bind     228..247
                     /label="BII(R)"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          228..247
                     /label="063_choi_pe_BIIR_PB_fwd"
                     /note="sequence: TTTACAGCGTGATGGAGCAG"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    286..289
                     /label="TTAA"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    290..302
                     /label="13 bp terminal repeat (TR)"
                     /ApEinfo_revcolor=#cd0532
                     /ApEinfo_fwdcolor=#cd0532
     misc_feature    290..352
                     /label="5'PB Terminal Repeat"
                     /ApEinfo_revcolor=#0080ff
                     /ApEinfo_fwdcolor=#0080ff
     misc_feature    303..333
                     /label="31 bp spacer"
                     /ApEinfo_revcolor=#0080c0
                     /ApEinfo_fwdcolor=#0080c0
     misc_feature    334..352
                     /label="19 bp Internal Repeat"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    528..759
                     /label="core Insulator"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     primer          744..773
                     /label="024_wc_pe_PB-5gibson"
                     /note="sequence: GGGGATACGGGGAAAAGGCCTCCACGGCCA"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    744..773
                     /label="5' gibson"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    773..773
                     /label="CMV7\promoter"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    782..789
                     /label="SbfI"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          complement(784..808)
                     /label="014_wc_tf_p7_amp_rev_nouse"
                     /note="sequence: CATTATATACCCTCTAGTGCCTGCA"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    790..843
                     /label="minimal promoter"
                     /ApEinfo_revcolor=#ff00ff
                     /ApEinfo_fwdcolor=#ff00ff
     misc_feature    815..844
                     /label="gibson"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     primer          829..888
                     /label="069_wc_tf_csy4_5'csy4hp_minP_fwd"
                     /note="sequence: GCAATCCGGTACTGTACCGGTactgccgtataggcagcgccaccatggaccactacctcg"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     protein_bind    850..866
                     /label="Csy4 site"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     regulatory      867..876
                     /label="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     CDS             873..1433
                     /label="Csy4"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     CDS             873..1463
                     /label="Translation 873-1463"
     primer          complement(1434..1463)
                     /label="002_wc_NLS_rev"
                     /note="sequence: tcagactttcctcttcttcttgggggatcc"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     CDS             1440..1460
                     /label="SV40 NLS"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    1464..1597
                     /label="Triplex modified Lu"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          complement(1637..1659)
                     /label="068_wc_tf_plsmP_rev"
                     /note="sequence: CGAAGTTATTAGGTCCCTCGACG"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    complement(1638..1659)
                     /label="pLSmP-ass-R-i# sits here"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    complement(1660..1699)
                     /label="3'BlastGibson"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer          complement(1660..1699)
                     /label="026_wc_pe_PB_3gibson_v2"
                     /note="sequence: TATTTGTAACCATTATAAGCTGCAATAAACAAGTTTAGTT"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    1665..1795
                     /label="SV40 polyA"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          complement(1672..1701)
                     /label="004_wc_SV40_rev"
                     /note="sequence: TTTATTTGTAACCATTATAAGCTGCAATAA"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    2044..2274
                     /label="core Insulator"
                     /ApEinfo_revcolor=#ff0080
                     /ApEinfo_fwdcolor=#ff0080
     misc_feature    2549..2567
                     /label="19 bp Internal Repeat(IR)"
                     /ApEinfo_revcolor=#ff0080
                     /ApEinfo_fwdcolor=#ff0080
     misc_feature    2549..2583
                     /label="3' PB Terminal Repeat"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    2568..2570
                     /label="3 bp spacer"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    2571..2583
                     /label="13 bp Terminal Repeat (TR)"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    2584..2587
                     /label="TTAA"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    complement(2790..2809)
                     /label="Potential primer mismatch"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer_bind     complement(3102..3121)
                     /label="pLac(F)"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          3243..3264
                     /label="103_am_join_rev1"
                     /note="sequence: CATTAATGAATCGGCCAACGCG"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    3275..3299
                     /label="5' gibson overhang"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    3303..3327
                     /label="3' gibson overhang"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    3476..4064
                     /label="ORI (mismatch with STAR by 1)"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer          complement(3556..3575)
                     /label="062_choi_pe_BR322Ori_rev"
                     /note="sequence: gggaaacgcctggtatcttt"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     CDS             complement(4238..5095)
                     /label="Amp"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
ORIGIN
        1 GTCCCATTCG CCATTCAGGC TGCGCAACTG TTGGGAAGGG CGATCGGTGC GGGCCTCTTC
       61 GCTATTACGC CAGCTGGCGA AAGGGGGATG TGCTGCAAGG CGATTAAGTT GGGTAACGCC
      121 AGGGTTTTCC CAGTCACGAC GTTGTAAAAC GACGGCCAGT GAGCGCGCCT CGTTCATTCA
      181 CGTTTTTGAA CCCGTGGAGG ACGGGCAGAC TCGCGGTGCA AATGTGTTTT ACAGCGTGAT
      241 GGAGCAGATG AAGATGCTCG ACACGCTGCA GAACACGCAG CTAGATTAAC CCTAGAAAGA
      301 TAATCATATT GTGACGTACG TTAAAGATAA TCATGTGTAA AATTGACGCA TGTGTTTTAT
      361 CGGTCTGTAT ATCGAGGTTT ATTTATTAAT TTGAATAGAT ATTAAGTTTT ATTATATTTA
      421 CACTTACATA CTAATAATAA ATTCAACAAA CAATTTATTT ATGTTTATTT ATTTATTAAA
      481 AAAAACAAAA ACTCAAAATT TCTTCTATAA AGTAACAAAA CTTTTATGAG GGACAGCCCC
      541 CCCCCAAAGC CCCCAGGGAT GTAATTACGT CCCTCCCCCG CTAGGGGGCA GCAGCGAGCC
      601 GCCCGGGGCT CCGCTCCGGT CCGGCGCTCC CCCCGCATCC CCGAGCCGGC AGCGTGCGGG
      661 GACAGCCCGG GCACGGGGAA GGTGGCACGG GATCGCTTTC CTCTGAACGC TTCTCGCTGC
      721 TCTTTGAGCC TGCAGACACC TGGGGGGATA CGGGGAAAAG GCCTCCACGG CCAGCTCTAG
      781 ACCTGCAGGC ACTAGAGGGT ATATAATGGA AGCTCGACTT CCAGCTTGGC AATCCGGTAC
      841 TGTACCGGTa ctgccgtata ggcagcgcca ccatggacca ctacctcgac attcgcttgc
      901 gaccggaccc ggaatttccc ccggcgcaac tcatgagcgt gctcttcggc aagctccacc
      961 aggccctggt ggcacagggc ggggacagga tcggcgtgag cttccccgac ctcgacgaaa
     1021 gccgctcccg gctgggcgag cgcctgcgca ttcatgcctc ggcggacgac cttcgtgccc
     1081 tgctcgcccg gccctggctg gaagggttgc gggaccatct gcaattcgga gaaccggcag
     1141 tcgtgcctca ccccacaccg taccgtcagg tcagtcgggt tcaggcgaaa agcaatccgg
     1201 aacgcctgcg gcggcggctc atgcgccggc acgatctgag tgaggaggag gctcggaaac
     1261 gcattcccga tacggtcgcg agagccttgg acctgccctt cgtcacgcta cgcagccaga
     1321 gcaccggaca gcacttccgt ctgttcatcc gccacgggcc gttgcaggtg acggcagagg
     1381 aaggaggatt cacctgttac gggttgagca aaggaggttt cgttccctgg ttcggatccc
     1441 ccaagaagaa gaggaaagtc tgatcagtag ggttgtaaag gtttttcttt tcctgagaaa
     1501 acaacctttt gttttctcag gttttgcttt ttggcctttc cctagcttta aaaaaaaaaa
     1561 agcaaaactc accgaggcag ttcgtatcgg tctgcgacac ttgagacgat taaATGGACA
     1621 GCAtgcgtct caAGTCCGTC GAGGGACCTA ATAACTTCGA ACTAAACTTG TTTATTGCAG
     1681 CTTATAATGG TTACAAATAA AGCAATAGCA TCACAAATTT CACAAATAAA GCATTTTTTT
     1741 CACTGCATTC TAGTTGTGGT TTGTCCAAAC TCATCAATGT ATCTTATCAT GTCTGGAATT
     1801 GACTCAAATG ATGTCAATTA GTCTATCAGA AGCTCATCTG GTCTCCCTTC CGGGGGACAA
     1861 GACATCCCTG TTTAATATTT AAACAGCAGT GTTCCCAAAC TGGGTTCTTA TATCCCTTGC
     1921 TCTGGTCAAC CAGGTTGCAG GGTTTCCTGT CCTCACAGGA ACGAAGTCCC TAAAGAAACA
     1981 GTGGCAGCCA GGTTTAGCCC CGGAATTGAC TGGATTCCTT TTTTAGGGCC CATTGGTATG
     2041 GCTTTTTCCC CGTATCCCCC CAGGTGTCTG CAGGCTCAAA GAGCAGCGAG AAGCGTTCAG
     2101 AGGAAAGCGA TCCCGTGCCA CCTTCCCCGT GCCCGGGCTG TCCCCGCACG CTGCCGGCTC
     2161 GGGGATGCGG GGGGAGCGCC GGACCGGAGC GGAGCCCCGG GCGGCTCGCT GCTGCCCCCT
     2221 AGCGGGGGAG GGACGTAATT ACATCCCTGG GGGCTTTGGG GGGGGGCTGT CCCTGATATC
     2281 TATAACAAGA AAATATATAT ATAATAAGTT ATCACGTAAG TAGAACATGA AATAACAATA
     2341 TAATTATCGT ATGAGTTAAA TCTTAAAAGT CACGTAAAAG ATAATCATGC GTCATTTTGA
     2401 CTCACGCGGT CGTTATAGTT CAAAATCAGT GACACTTACC GCATTGACAA GCACGCCTCA
     2461 CGGGAGCTCC AAGCGGCGAC TGAGATGTCC TAAATGCACA GCGACGGATT CGCGCTATTT
     2521 AGAAAGAGAG AGCAATATTT CAAGAATGCA TGCGTCAATT TTACGCAGAC TATCTTTCTA
     2581 GGGTTAATCT AGCTGCATCA GGATCATATC GTCGGGTCTT TTTTCCGGCT CAGTCATCGC
     2641 CCAAGCTGGC GCTATCTGGG CATCGGGGAG GAAGAAGCCC GTGCCTTTTC CCGCGAGGTT
     2701 GAAGCGGCAT GGAAAGAGTT TGCCGAGGAT GACTGCTGCT GCATTGACGT TGAGCGAAAA
     2761 CGCACGTTTA CCATGATGAT TCGGGAAGGT GTGGCCATGC ACGCCTTTAA CGGTGAACTG
     2821 TTCGTTCAGG CCACCTGGGA TACCAGTTCG TCGCGGCTTT TCCGGACACA GTTCCGGATG
     2881 GTCAGCCCGA AGCGCATCAG CAACCCGAAC AATACCGGCG ACAGCCGGAA CTGCCGTGCC
     2941 GGTGTGCAGA TTAATGACAG CGGTGCGGCG CTGGGATATT ACGTCAGCGA GGACGGGTAT
     3001 CCTGGCTGGA TGCCGCAGAA ATGGACATGG ATACCCCGTG AGTTACCCGG CGGGCGCGCT
     3061 TGGCGTAATC ATGGTCATAG CTGTTTCCTG TGTGAAATTG TTATCCGCTC ACAATTCCAC
     3121 ACAACATACG AGCCGGAAGC ATAAAGTGTA AAGCCTGGGG TGCCTAATGA GTGAGCTAAC
     3181 TCACATTAAT TGCGTTGCGC TCACTGCCCG CTTTCCAGTC GGGAAACCTG TCGTGCCAGC
     3241 TGCATTAATG AATCGGCCAA CGCGCGGGGA GAGGCGGTTT GCGTATTGGG CGCTCTTCCG
     3301 CTTCCTCGCT CACTGACTCG CTGCGCTCGG TCGTTCGGCT GCGGCGAGCG GTATCAGCTC
     3361 ACTCAAAGGC GGTAATACGG TTATCCACAG AATCAGGGGA TAACGCAGGA AAGAACATGT
     3421 GAGCAAAAGG CCAGCAAAAG GCCAGGAACC GTAAAAAGGC CGCGTTGCTG GCGTTTTTCC
     3481 ATAGGCTCCG CCCCCCTGAC GAGCATCACA AAAATCGACG CTCAAGTCAG AGGTGGCGAA
     3541 ACCCGACAGG ACTATAAAGA TACCAGGCGT TTCCCCCTGG AAGCTCCCTC GTGCGCTCTC
     3601 CTGTTCCGAC CCTGCCGCTT ACCGGATACC TGTCCGCCTT TCTCCCTTCG GGAAGCGTGG
     3661 CGCTTTCTCA TAGCTCACGC TGTAGGTATC TCAGTTCGGT GTAGGTCGTT CGCTCCAAGC
     3721 TGGGCTGTGT GCACGAACCC CCCGTTCAGC CCGACCGCTG CGCCTTATCC GGTAACTATC
     3781 GTCTTGAGTC CAACCCGGTA AGACACGACT TATCGCCACT GGCAGCAGCC ACTGGTAACA
     3841 GGATTAGCAG AGCGAGGTAT GTAGGCGGTG CTACAGAGTT CTTGAAGTGG TGGCCTAACT
     3901 ACGGCTACAC TAGAAGGACA GTATTTGGTA TCTGCGCTCT GCTGAAGCCA GTTACCTTCG
     3961 GAAAAAGAGT TGGTAGCTCT TGATCCGGCA AACAAACCAC CGCTGGTAGC GGTGGTTTTT
     4021 TTGTTTGCAA GCAGCAGATT ACGCGCAGAA AAAAAGGATC TCAAGAAGAT CCTTTGATCT
     4081 TTTCTACGGG GTCTGACGCT CAGTGGAACG AAAACTCACG TTAAGGGATT TTGGTCATGA
     4141 GATTATCAAA AAGGATCTTC ACCTAGATCC TTTTAAATTA AAAATGAAGT TTTAAATCAA
     4201 TCTAAAGTAT ATATGAGTAA ACTTGGTCTG ACAGTTACCA ATGCTTAATC AGTGAGGCAC
     4261 CTATCTCAGC GATCTGTCTA TTTCGTTCAT CCATAGTTGC CTGACTCCCC GTCGTGTAGA
     4321 TAACTACGAT ACGGGAGGGC TTACCATCTG GCCCCAGTGC TGCAATGATA CCGCGAGACC
     4381 CACGCTCACC GGCTCCAGAT TTATCAGCAA TAAACCAGCC AGCCGGAAGG GCCGAGCGCA
     4441 GAAGTGGTCC TGCAACTTTA TCCGCCTCCA TCCAGTCTAT TAATTGTTGC CGGGAAGCTA
     4501 GAGTAAGTAG TTCGCCAGTT AATAGTTTGC GCAACGTTGT TGCCATTGCT ACAGGCATCG
     4561 TGGTGTCACG CTCGTCGTTT GGTATGGCTT CATTCAGCTC CGGTTCCCAA CGATCAAGGC
     4621 GAGTTACATG ATCCCCCATG TTGTGCAAAA AAGCGGTTAG CTCCTTCGGT CCTCCGATCG
     4681 TTGTCAGAAG TAAGTTGGCC GCAGTGTTAT CACTCATGGT TATGGCAGCA CTGCATAATT
     4741 CTCTTACTGT CATGCCATCC GTAAGATGCT TTTCTGTGAC TGGTGAGTAC TCAACCAAGT
     4801 CATTCTGAGA ATAGTGTATG CGGCGACCGA GTTGCTCTTG CCCGGCGTCA ATACGGGATA
     4861 ATACCGCGCC ACATAGCAGA ACTTTAAAAG TGCTCATCAT TGGAAAACGT TCTTCGGGGC
     4921 GAAAACTCTC AAGGATCTTA CCGCTGTTGA GATCCAGTTC GATGTAACCC ACTCGTGCAC
     4981 CCAACTGATC TTCAGCATCT TTTACTTTCA CCAGCGTTTC TGGGTGAGCA AAAACAGGAA
     5041 GGCAAAATGC CGCAAAAAAG GGAATAAGGG CGACACGGAA ATGTTGAATA CTCATACTCT
     5101 TCCTTTTTCA ATATTATTGA AGCATTTATC AGGGTTATTG TCTCATGAGC GGATACATAT
     5161 TTGAATGTAT TTAGAAAAAT AAACAAATAG GGGTTCCGCG CACATTTCCC CGAAAAGTGC
     5221 CACCTAAATT GTAAGCGTTA ATATTTTGTT AAAATTCGCG TTAAATTTTT GTTAAATCAG
     5281 CTCATTTTTT AACCAATAGG CCGAAATCGG CAAAATCCCT TATAAATCAA AAGAATAGAC
     5341 CGAGATAGGG TTGAGTGTTG TTCCAGTTTG GAACAAGAGT CCACTATTAA AGAACGTGGA
     5401 CTCCAACGTC AAAGGGCGAA AAACCGTCTA TCAGGGCGAT GGCCCACTAC GTGAACCATC
     5461 ACCCTAATCA AGTTTTTTGG GGTCGAGGTG CCGTAAAGCA CTAAATCGGA ACCCTAAAGG
     5521 GAGCCCCCGA TTTAGAGCTT GACGGGGAAA GCCGGCGAAC GTGGCGAGAA AGGAAGGGAA
     5581 GAAAGCGAAA GGAGCGGGCG CTAGGGCGCT GGCAAGTGTA GCGGTCACGC TGCGCGTAAC
     5641 CACCACACCC GCCGCGCTTA ATGCGCCGCT ACAGGGCGC
//