LOCUS       Intron-Tagging-mScarlet 5096 bp ds-DNA     circular     29-JAN-2024
DEFINITION  .
FEATURES             Location/Qualifiers
     misc_feature    2291..2325
                     /label="attB"
     primer          2327..2357
                     /label="frame1_mscarlet_fwd"
                     /note="sequence: CTAGTGAATTCAGATCTGATcctgcgttatc"
     primer          2327..2365
                     /label="minicircle_fwd"
                     /note="sequence: CTAGTGAATTCAGATCTGATcctgcgttatcccctggag"
     primer          2327..2435
                     /label="frame1_leftarm1"
                     /note="sequence: CTAGTGAATTCAGATCTGATcctgcgttatcccctggagatcgagtgccgcatcacCGGCTATTGGTCTTACTGACATCCACTTTGCCTTTCTCTCCACAGggggtggc"
     misc_feature    2363..2382
                     /label="generic sgRNA target"
     misc_feature    2383..2385
                     /label="PAM"
     misc_feature    2386..2427
                     /label="splice acceptor"
     misc_feature    2401..2401
                     /label="BP A"
     primer          complement(2404..2515)
                     /label="frame1_leftarm2"
                     /note="sequence: TTGATCACTGCCTCGCCCTTGCTCACggatccaccaccgccagatccgccgccaccggacccaccaccgcccgagccaccgccaccccCTGTGGAGAGAAAGGCAAAGTGGA"
     misc_feature    2428..2429
                     /label=+2
     misc_feature    2430..2489
                     /label="(GGGGS)x4 linker"
     CDS             2430..2489
                     /label="Translation 2430-2489"
                     /translation="GGGGSGGGGSGGGGSGGGGS"
     primer          complement(2489..2504)
                     /label="integration_rev_second"
                     /note="sequence: ctcgcccttgctcacg"
     misc_feature    2490..3182
                     /label="mScarlet-I"
     CDS             2490..3182
                     /label="Translation 2490-3182"
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGGAVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKMALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHSTGGMDELYK"
     primer          2491..2506
                     /label="mScarlet_fwd"
                     /note="sequence: TGAGCAAGGGCGAGGC"
     primer          2491..2508
                     /label="mScarlet_fwd2"
                     /note="sequence: TGAGCAAGGGCGAGGCAG"
     primer          complement(2496..2515)
                     /label="frame1_mscarlet_rev"
                     /note="sequence: TTGATCACTGCCTCGCCCTT"
     primer          complement(2498..2515)
                     /label="mScarlet_left arm_rev"
                     /note="sequence: TTGATCACTGCCTCGCCC"
     primer          3157..3173
                     /label="mScarlet_ right arm fwd"
                     /note="sequence: CCGGCGGCATGGACGAG"
     primer          3157..3174
                     /label="frame1_mscarlet_F"
                     /note="sequence: CCGGCGGCATGGACGAGC"
     primer          complement(3157..3181)
                     /label="mScarlet_rev"
                     /note="sequence: TTGTACAGCTCGTCCATGCCGCCGG"
     primer          3157..3273
                     /label="frame1_rightarm1"
                     /note="sequence: CCGGCGGCATGGACGAGCTGTACAAGggcggcggtggatccggtggcggcggatctggcggtggaggttcgggtggcggtgggtcgggcggagGTAAGTATCAATTAGAAGACGAAT"
     primer          complement(3162..3181)
                     /label="mScarlet_rev2"
                     /note="sequence: TTGTACAGCTCGTCCATGCC"
     misc_feature    3183..3242
                     /label="(GGGGS)x4 linker"
     CDS             3183..3245
                     /label="Translation 3183-3245"
                     /translation="GGGGSGGGGSGGGGSGGGGSG"
     primer          complement(3240..3295)
                     /label="frame1_rightarm2"
                     /note="sequence: ATGGGTCGACTCTAGAGATGGAATTCGTCTTCTAATTGATACTTACctccgcccga"
     CDS             3246..3248
                     /label="Translation 3246-3248"
                     /translation="G"
     misc_feature    3249..3249
                     /label=+1
     misc_feature    3250..3269
                     /label="splice donor"
     primer          complement(3268..3295)
                     /label="frame1_mscarlet_R"
                     /note="sequence: ATGGGTCGACTCTAGAGATGGAATTCGT"
     misc_feature    3276..3276
                     /label="PAM"
     misc_feature    3306..3342
                     /label="attP"
     misc_feature    complement(3430..4224)
                     /label="KanR"
     misc_feature    5096..588
                     /label="ori"
ORIGIN
        1 TTGAGATCCT TTTTTTCTGC GCGTAATCTG CTGCTTGCAA ACAAAAAAAC CACCGCTACC
       61 AGCGGTGGTT TGTTTGCCGG ATCAAGAGCT ACCAACTCTT TTTCCGAAGG TAACTGGCTT
      121 CAGCAGAGCG CAGATACCAA ATACTGTCCT TCTAGTGTAG CCGTAGTTAG GCCACCACTT
      181 CAAGAACTCT GTAGCACCGC CTACATACCT CGCTCTGCTA ATCCTGTTAC CAGTGGCTGC
      241 TGCCAGTGGC GATAAGTCGT GTCTTACCGG GTTGGACTCA AGACGATAGT TACCGGATAA
      301 GGCGCAGCGG TCGGGCTGAA CGGGGGGTTC GTGCACACAG CCCAGCTTGG AGCGAACGAC
      361 CTACACCGAA CTGAGATACC TACAGCGTGA GCTATGAGAA AGCGCCACGC TTCCCGAAGG
      421 GAGAAAGGCG GACAGGTATC CGGTAAGCGG CAGGGTCGGA ACAGGAGAGC GCACGAGGGA
      481 GCTTCCAGGG GGAAACGCCT GGTATCTTTA TAGTCCTGTC GGGTTTCGCC ACCTCTGACT
      541 TGAGCGTCGA TTTTTGTGAT GCTCGTCAGG GGGGCGGAGC CTATGGAAAA ACGCCAGCAA
      601 CGCGGCCTTT TTACGGTTCC TGGCCTTTTG CTGGCCTTTT GCTCACATGT TCTTTCCTGC
      661 GTTATCCCCT GATTCTGTGG ATAACCGTAT TACCGCCTTT GAGTGAGCTG ATACCGCTCG
      721 CCGCAGCCGA ACGACCGAGC GCAGCGAGTC AGTGAGCGAG GAAGCGGAAG AGCGCCTGAT
      781 GCGGTATTTT CTCCTTACGC ATCTGTGCGG TATTTCACAC CGCATATGGT GCACTCTCAG
      841 TACAATCTGC TCTGATGCCG CATAGTTAAG CCAGTATACA CTCCGCTATC GCTACGTGAC
      901 TGGGTCATGG CTGCGCCCCG ACACCCGCCA ACACCCGCTG ACGCGCCCTG ACGGGCTTGT
      961 CTGCTCCCGG CATCCGCTTA CAGACAAGCT GTGACCGTCT CCGGGAGCTG CATGTGTCAG
     1021 AGGTTTTCAC CGTCATCACC GAAACGCGCG AGGCAGCAGA TCAATTCGCG CGCGAAGGCG
     1081 AAGCGGCATG CATAATGTGC CTGTCAAATG GACGAAGCAG GGATTCTGCA AACCCTATGC
     1141 TACTCCGTCA AGCCGTCAAT TGTCTGATTC GTTACCAATT ATGACAACTT GACGGCTACA
     1201 TCATTCACTT TTTCTTCACA ACCGGCACGG AACTCGCTCG GGCTGGCCCC GGTGCATTTT
     1261 TTAAATACCC GCGAGAAATA GAGTTGATCG TCAAAACCAA CATTGCGACC GACGGTGGCG
     1321 ATAGGCATCC GGGTGGTGCT CAAAAGCAGC TTCGCCTGGC TGATACGTTG GTCCTCGCGC
     1381 CAGCTTAAGA CGCTAATCCC TAACTGCTGG CGGAAAAGAT GTGACAGACG CGACGGCGAC
     1441 AAGCAAACAT GCTGTGCGAC GCTGGCGATT CTTCTCTCAT CCGCCAAAAC AGCCAAGCTG
     1501 GAGACCGTTT GACATTACCC TGTTATCCCT AGATACATTA CCCTGTTATC CCAGATGACA
     1561 TACCCTGTTA TCCCTAGATG ACATTACCCT GTTATCCCAG ATGACATTAC CCTGTTATCC
     1621 CTAGATACAT TACCCTGTTA TCCCAGATGA CATACCCTGT TATCCCTAGA TGACATTACC
     1681 CTGTTATCCC AGATGACATT ACCCTGTTAT CCCTAGATAC ATTACCCTGT TATCCCAGAT
     1741 GACATACCCT GTTATCCCTA GATGACATTA CCCTGTTATC CCAGATGACA TTACCCTGTT
     1801 ATCCCTAGAT ACATTACCCT GTTATCCCAG ATGACATACC CTGTTATCCC TAGATGACAT
     1861 TACCCTGTTA TCCCAGATGA CATTACCCTG TTATCCCTAG ATACATTACC CTGTTATCCC
     1921 AGATGACATA CCCTGTTATC CCTAGATGAC ATTACCCTGT TATCCCAGAT GACATTACCC
     1981 TGTTATCCCT AGATACATTA CCCTGTTATC CCAGATGACA TACCCTGTTA TCCCTAGATG
     2041 ACATTACCCT GTTATCCCAG ATGACATTAC CCTGTTATCC CTAGATACAT TACCCTGTTA
     2101 TCCCAGATGA CATACCCTGT TATCCCTAGA TGACATTACC CTGTTATCCC AGATGACATT
     2161 ACCCTGTTAT CCCTAGATAC ATTACCCTGT TATCCCAGAT GACATACCCT GTTATCCCTA
     2221 GATGACATTA CCCTGTTATC CCAGATAAAC TCAATGATGA TGATGATGAT GGTCGAGACT
     2281 CAGCGGCCGC GGTGCCAGGG CGTGCCCTTG GGCTCCCCGG GCGCGACTAG TGAATTCAGA
     2341 TCTGATcctg cgttatcccc tggagatcga gtgccgcatc acCGGCTATT GGTCTTACTG
     2401 ACATCCACTT TGCCTTTCTC TCCACAGggg gtggcggtgg ctcgggcggt ggtgggtccg
     2461 gtggcggcgg atctggcggt ggtggatccG TGAGCAAGGG CGAGGCAGTG ATCAAGGAGT
     2521 TCATGCGGTT CAAGGTGCAC ATGGAGGGCT CCATGAACGG CCACGAGTTC GAGATCGAGG
     2581 GCGAGGGCGA GGGCCGCCCC TACGAGGGCA CCCAGACCGC CAAGCTGAAG GTGACCAAGG
     2641 GTGGCCCCCT GCCCTTCTCC TGGGACATCC TGTCCCCTCA GTTCATGTAC GGCTCCAGGG
     2701 CCTTCATCAA GCACCCCGCC GACATCCCCG ACTACTATAA GCAGTCCTTC CCCGAGGGCT
     2761 TCAAGTGGGA GCGCGTGATG AACTTCGAGG ACGGCGGCGC CGTGACCGTG ACCCAGGACA
     2821 CCTCCCTGGA GGACGGCACC CTGATCTACA AGGTGAAGCT CCGCGGCACC AACTTCCCTC
     2881 CTGACGGCCC CGTAATGCAG AAGAAGACAA TGGGCTGGGA AGCGTCCACC GAGCGGTTGT
     2941 ACCCCGAGGA CGGCGTGCTG AAGGGCGACA TTAAGATGGC CCTGCGCCTG AAGGACGGCG
     3001 GCCGCTACCT GGCGGACTTC AAGACCACCT ACAAGGCCAA GAAGCCCGTG CAGATGCCCG
     3061 GCGCCTACAA CGTCGACCGC AAGTTGGACA TCACCTCCCA CAACGAGGAC TACACCGTGG
     3121 TGGAACAGTA CGAACGCTCC GAGGGCCGCC ACTCCACCGG CGGCATGGAC GAGCTGTACA
     3181 AGggcggcgg tggatccggt ggcggcggat ctggcggtgg aggttcgggt ggcggtgggt
     3241 cgggcggagG TAAGTATCAA TTAGAAGACG AATTCCATCT CTAGAGTCGA CCCATGGGGG
     3301 CCCGCCCCAA CTGGGGTAAC CTTTGAGTTC TCTCAGTTGG GGGTAATCAG CATCATGATG
     3361 TGGTACCACA TCATGATGCT GATTATAAGA ATGCGGCCGC CACACTCTAG TGGATCTCGA
     3421 GTTAATAATT CAGAAGAACT CGTCAAGAAG GCGATAGAAG GCGATGCGCT GCGAATCGGG
     3481 AGCGGCGATA CCGTAAAGCA CGAGGAAGCG GTCAGCCCAT TCGCCGCCAA GCTCTTCAGC
     3541 AATATCACGG GTAGCCAACG CTATGTCCTG ATAGCGGTCC GCCACACCCA GCCGGCCACA
     3601 GTCGATGAAT CCAGAAAAGC GGCCATTTTC CACCATGATA TTCGGCAAGC AGGCATCGCC
     3661 ATGGGTCACG ACGAGATCCT CGCCGTCGGG CATGCTCGCC TTGAGCCTGG CGAACAGTTC
     3721 GGCTGGCGCG AGCCCCTGAT GCTCTTCGTC CAGATCATCC TGATCGACAA GACCGGCTTC
     3781 CATCCGAGTA CGTGCTCGCT CGATGCGATG TTTCGCTTGG TGGTCGAATG GGCAGGTAGC
     3841 CGGATCAAGC GTATGCAGCC GCCGCATTGC ATCAGCCATG ATGGATACTT TCTCGGCAGG
     3901 AGCAAGGTGA GATGACAGGA GATCCTGCCC CGGCACTTCG CCCAATAGCA GCCAGTCCCT
     3961 TCCCGCTTCA GTGACAACGT CGAGCACAGC TGCGCAAGGA ACGCCCGTCG TGGCCAGCCA
     4021 CGATAGCCGC GCTGCCTCGT CTTGCAGTTC ATTCAGGGCA CCGGACAGGT CGGTCTTGAC
     4081 AAAAAGAACC GGGCGCCCCT GCGCTGACAG CCGGAACACG GCGGCATCAG AGCAGCCGAT
     4141 TGTCTGTTGT GCCCAGTCAT AGCCGAATAG CCTCTCCACC CAAGCGGCCG GAGAACCTGC
     4201 GTGCAATCCA TCTTGTTCAA TCATGCGAAA CGATCCTCAT CCTGTCTCTT GATCAGAGCT
     4261 TGATCCCCTG CGCCATCAGA TCCTTGGCGG CAAGAAAGCC ATCCAGTTTA CTTTGCAGGG
     4321 CTTCCCAACC TTACCAGAGG GCGCCCCAGC TGGCAATTCC GGTTCGCTTG CTGTCCATAA
     4381 AACCGCCCAG TCTAGCTATC GCCATGTAAG CCCACTGCAA GCTACCTGCT TTCTCTTTGC
     4441 GCTTGCGTTT TCCCTTGTCC AGATAGCCCA GTAGCTGACA TTCATCCGGG GTCAGCACCG
     4501 TTTCTGCGGA CTGGCTTTCT ACGCGCTCGA GGgggGgccA AACGGTCTCC AGCTTGGCTG
     4561 TTTTGGCGGA TGAGAGAAGA TTTTCAGCCT GATACAGATT AAATCAGAAC GCAGAAGCGG
     4621 TCTGATAAAA CAGAATTTGC CTGGCGGCAG TAGCGCGGTG GTCCCACCTG ACCCCATGCC
     4681 GAACTCAGAA GTGAAACGCC GTAGCGCCGA TGGTAGTGTG GGGTCTCCCC ATGCGAGAGT
     4741 AGGGAACTGC CAGGCATCAA ATAAAACGAA AGGCTCAGTC GAAAGACTGG GCCTTTCGTT
     4801 TTATCTGTTG TTTGTCGGTG AACGCTCTCC TGAGTAGGAC AAATCCGCCG GGAGCGGATT
     4861 TGAACGTTGC GAAGCAACGG CCCGGAGGGT GGCGGGCAGG ACGCCCGCCA TAAACTGCCA
     4921 GGCATCAAAT TAAGCAGAAG GCCATCCTGA CGGATGGCCT TTTTGCGTTT CTACAAACTC
     4981 TTTTGTTTAT TTTTCTAAAT ACATTCAAAT ATGTATCCGC TCATGACCAA AATCCCTTAA
     5041 CGTGAGTTTT CGTTCCACTG AGCGTCAGAC CCCGTAGAAA AGATCAAAGG ATCTTC
//