LOCUS       pBbAk-W4-loxP-TT-loxP-m 5200 bp ds-DNA     circular     10-OCT-2024
DEFINITION  .
FEATURES             Location/Qualifiers
     misc_feature    93..107
                     /label="USR(Upstream)"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     promoter        93..163
                     /label="T7A1 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     unsure          108..113
                     /label="-35 site"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    111..112
                     /label="AC"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    114..130
                     /label="Core"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     unsure          131..136
                     /label="-10 site"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    133..134
                     /label="TA"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    137..163
                     /label="DSR(Downstream)"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     unsure          144..144
                     /label="+1 site from Brunner, M. etc al. 1987"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_recomb     164..197
                     /label="LoxP"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    237..365
                     /label="dbl term"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_recomb     368..401
                     /label="LoxP"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     RBS             402..421
                     /label="RBS"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    complement(422..436)
                     /label="misc_feature"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    437..1138
                     /label="mCherry"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer          complement(507..529)
                     /label="seq_mCherry_rev1"
                     /note="sequence: ctcgaactcgtgaccgttaacag"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    1139..1144
                     /label="Stop"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    1188..1316
                     /label="dbl term"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer          complement(1302..1322)
                     /label="FJ-0097"
                     /note="sequence: tacggaaggatctgacctaggtataaacgcagaaag"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     rep_origin      complement(1323..2034)
                     /label="p15a"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          complement(1467..1486)
                     /label="oES105 [Seq: JBEI-6409 ispA Insert] REV"
                     /note="sequence: cggagcctatggaaaaacgg"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          1971..1990
                     /label="FJ-0011"
                     /note="sequence: tttcgttttcagagcaagag"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer          complement(2041..2066)
                     /label="FJ-0024"
                     /note="sequence: AGGTAGGGTCTCCAGattgcactgaaatctagaaatatttt"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          2067..2084
                     /label="FJ-0021"
                     /note="sequence: AGGTAGGGTCTCGatCTGACAGCTAGCTCAGTC"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    2067..2101
                     /label="Constitutive promoter J23108"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    2067..3760
                     /label="PCB Production Region"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     RBS             2116..2129
                     /label="RBS pprotet"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    2118..2129
                     /label="B0034"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             2136..2858
                     /label="ho1 (RBS 80.077)"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     RBS             2859..2872
                     /label="RBS pprotet"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    2861..2872
                     /label="B0034"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             2879..3625
                     /label="pcyA"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    2923..2923
                     /label="Silent mutation to C"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    3632..3760
                     /label="Terminator B0015"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          complement(3742..3760)
                     /label="FJ-0022"
                     /note="sequence: AGGTAGGGTCTCGctaTATAAACGCAGAAAGGCCC"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer          3761..3778
                     /label="FJ-0023"
                     /note="sequence: AGGTAGGGTCTCCAtagcacctgaagtcagcc"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     terminator      3804..3909
                     /label="TO"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    complement(3935..4729)
                     /label="Kan/neoR"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer          complement(3993..4012)
                     /label="FJ-0012"
                     /note="sequence: ttcctcgtgctttacggtat"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    4894..4913
                     /label="Ligation"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer          5181..5198
                     /label="seq_E5a-Cre"
                     /note="sequence: cccgatcttccccatcgg"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer_bind     5197..15
                     /label="Seq-F-X0"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
ORIGIN
        1 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
       61 ccgaagttcc tattatcacg aggccctttg agTTATCAAA AAGAGTATTG CATTAAAGTC
      121 TAACCTATAG GAATCTTACA GCCATCGAGA GGGACACGGC GAAataactt cgtatagcat
      181 acattatacg aagttatcca ggcatcaaat aaggatccaa actcgagtaa ggatctccag
      241 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
      301 gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt
      361 ttataccata acttcgtata gcatacatta tacgaagtta tTTTAAGAAG GAGATATACA
      421 Tatgactagc aaaagatcca agggcgagga ggataacatg gctatcatta aagagttcat
      481 gcgcttcaaa gttcacatgg agggttctgt taacggtcac gagttcgaga tcgaaggcga
      541 aggcgagggc cgtccgtatg aaggcaccca gaccgccaaa ctgaaagtga ctaaaggcgg
      601 cccgctgcct tttgcgtggg acatcctgag cccgcaattt atgtacggtt ctaaagcgta
      661 tgttaaacac ccagcggata tcccggacta tctgaagctg tcttttccgg aaggtttcaa
      721 gtgggaacgc gtaatgaatt ttgaagatgg tggtgtcgtg accgtcactc aggactcctc
      781 cctgcaagat ggcgagttca tctataaagt taaactgcgt ggtactaatt ttccatctga
      841 tggcccggtg atgcagaaaa agacgatggg ttgggaggcg tctagcgaac gcatgtatcc
      901 ggaagatggt gcgctgaaag gcgaaattaa acagcgcctg aaactgaaag atggcggcca
      961 ttatgacgct gaagtgaaaa ccacgtacaa agccaagaaa cctgtgcagc tgcctggcgc
     1021 gtacaatgtg aatattaaac tggacatcac ctctcataat gaagattata cgatcgtaga
     1081 gcaatatgag cgcgcggagg gtcgtcattc taccggtggc atggatgagc tgtacaaata
     1141 atagGGATCT CCAGGCATCA AATAAGGATC CAAACTCGAG Tggatctcca ggcatcaaat
     1201 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1261 cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataccta
     1321 gggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
     1381 gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac
     1441 agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc
     1501 atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc
     1561 aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg
     1621 gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg
     1681 taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc
     1741 gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg
     1801 gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg
     1861 ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa
     1921 agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc
     1981 agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat
     2041 aaaatatttc tagatttcag tgcaatCTGA CAGCTAGCTC AGTCCTAGGT ATAATGCTAG
     2101 CTCACACAGA ATTCATTAAA GAGGAGAAAG GTACCATGAG TGTCAACTTA GCTTCCCAGT
     2161 TGCGGGAAGG GACGAAAAAA TCCCACTCCA TGGCGGAGAA CGTCGGCTTT GTCAAATGCT
     2221 TCCTCAAGGG CGTTGTCGAG AAAAATTCCT ACCGTAAGCT GGTTGGCAAT CTCTACTTTG
     2281 TCTACAGTGC CATGGAAGAG GAAATGGCAA AATTTAAGGA CCATCCCATC CTCAGCCACA
     2341 TTTACTTCCC CGAACTCAAC CGCAAACAAA GCCTAGAGCA AGACCTGCAA TTCTATTACG
     2401 GCTCCAACTG GCGGCAAGAA GTGAAAATTT CTGCCGCTGG CCAAGCCTAT GTGGACCGAG
     2461 TCCGGCAAGT GGCCGCTACG GCCCCTGAAT TGTTGGTGGC CCATTCCTAC ACCCGTTACC
     2521 TGGGGGATCT TTCCGGCGGT CAAATTCTCA AGAAAATTGC CCAAAATGCC ATGAATCTCC
     2581 ACGATGGTGG CACAGCTTTC TATGAATTTG CCGACATTGA TGACGAAAAG GCTTTTAAAA
     2641 ATACCTACCG TCAAGCTATG AATGATCTGC CCATTGACCA AGCCACCGCC GAACGGATTG
     2701 TGGATGAAGC CAATGACGCC TTTGCCATGA ACATGAAAAT GTTCAACGAA CTTGAAGGCA
     2761 ACCTGATCAA GGCGATCGGC ATTATGGTGT TCAACAGCCT CACCCGTCGC CGCAGTCAAG
     2821 GCAGCACCGA AGTTGGCCTC GCCACCTCCG AAGGCTAGTT AAAGAGGAGA AAGGATCCAT
     2881 GGCCGTCACT GATTTAAGTT TGACCAATTC TTCCCTGATG CCTACGTTGA ACCCGATGAT
     2941 TCAACAGTTG GCCCTGGCGA TCGCCGCTAG TTGGCAAAGT TTACCCCTCA AGCCCTATCA
     3001 ATTGCCGGAG GATTTGGGCT ACGTAGAAGG CCGCCTGGAA GGGGAAAAGT TAGTGATTGA
     3061 AAATCGGTGC TACCAAACGC CCCAGTTTCG CAAAATGCAT TTGGAGTTGG CCAAGGTGGG
     3121 CAAAGGGTTG GATATTCTCC ACTGTGTAAT GTTTCCTGAG CCTTTATACG GTCTACCTTT
     3181 GTTTGGCTGT GACATTGTGG CCGGCCCCGG TGGAGTAAGT GCGGCTATTG CGGATCTATC
     3241 CCCCACCCAA AGCGATCGCC AATTGCCCGC AGCGTACCAA AAATCATTGG CAGAGCTAGG
     3301 CCAGCCAGAA TTTGAGCAAC AACGGGAATT GCCCCCCTGG GGAGAAATAT TTTCTGAATA
     3361 TTGTTTATTC ATCCGTCCCA GCAATGTCAC TGAAGAAGAA AGATTTGTAC AAAGGGTAGT
     3421 GGACTTTTTG CAAATTCATT GTCACCAATC CATCGTTGCC GAACCCTTGT CTGAAGCTCA
     3481 AACTTTGGAG CACCGTCAGG GGCAAATTCA TTACTGCCAA CAACAACAGA AAAATGATAA
     3541 AACCCGTCGG GTACTGGAAA AAGCTTTTGG GGAAGCTTGG GCGGAACGGT ATATGAGCCA
     3601 AGTCTTATTT GATGTTATCC AATAATCTAG ACCAGGCATC AAATAAAACG AAAGGCTCAG
     3661 TCGAAAGACT GGGCCTTTCG TTTTATCTGT TGTTTGTCGG TGAACGCTCT CTACTAGAGT
     3721 CACACTGGCT CACCTTCGGG TGGGCCTTTC TGCGTTTATA tagcacctga agtcagcccc
     3781 atacgatata agttgttact agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa
     3841 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
     3901 agtccaagcg agctctcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat
     3961 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
     4021 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
     4081 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
     4141 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc
     4201 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
     4261 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
     4321 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
     4381 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
     4441 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
     4501 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca
     4561 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
     4621 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
     4681 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
     4741 ctcatcctgt ctcttgatca gatcatgatc ccctgcgcca tcagatcctt ggcggcaaga
     4801 aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca
     4861 attccgacgt cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgcattcacc
     4921 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     4981 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc
     5041 cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat
     5101 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
     5161 catgagcccg aagtggcgag cccgatcttc cccatcggtg
//