LOCUS       3'cassette_deltaH2B_FS_ 5276 bp ds-DNA     circular     20-JUL-2022
DEFINITION  .
FEATURES             Location/Qualifiers
     source          1..2
                     /label="source"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     primer_bind     536..537
                     /label="M13-fwd"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     promoter        583..794
                     /label="EF-1-alpha core promoter"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          complement(629..648)
                     /label="RG661"
                     /note="sequence: caccggatcaattgccgacc"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          631..650
                     /label="RG632"
                     /note="sequence: tcggcaattgatccggtgcc"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     primer          740..759
                     /label="RG662"
                     /note="sequence: tgcagtagtcgccgtgaacg"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     primer          803..850
                     /label="AV490"
                     /note="sequence: ctagagcgctcgcccgccccacgagagATGaaaaagcctgaactcacc"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          complement(803..850)
                     /label="AV491"
                     /note="sequence: ggtgagttcaggctttttCATctctcgtggggcgggcgagcgctctag"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          complement(812..832)
                     /label="AV544"
                     /note="sequence: CATctctcgtggggcgggcga"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     CDS             830..1852
                     /label="HygroR"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     CDS             830..2392
                     /label="Translation 830-2392"
     primer          851..870
                     /label="RG664"
                     /note="sequence: gcgacgtctgtcgagaagtt"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer          complement(851..870)
                     /label="RG615"
                     /note="sequence: aacttctcgacagacgtcgc"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     primer          865..886
                     /label="RG697"
                     /note="sequence: gaagtttctgatcgaaaagttc"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     primer          complement(1640..1661)
                     /label="RG698"
                     /note="sequence: agctctgatagagttggtcaag"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          complement(1833..1852)
                     /label="RG665"
                     /note="sequence: ttcctttgccctcggacgag"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    1853..1915
                     /label="T2A misc_feature"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     primer          1892..1911
                     /label="RG633"
                     /note="sequence: GACGTCGAGGAGAATCCTGG"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     primer          complement(1892..1941)
                     /label="AV496"
                     /note="sequence: ggagcagacttcgctggctctggcatTGGGCCAGGATTCTCCTCGACGTC"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     primer          1894..1948
                     /label="AV497"
                     /note="sequence: caccaagtacaccagcgctaagGGGGGCGGGGTGAGCAAGGGCGAGGAGCTGTTC"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          complement(1895..1915)
                     /label="AV538"
                     /note="sequence: TGGGCCAGGATTCTCCTCGAC"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer          1916..1938
                     /label="AV539"
                     /note="sequence: GGGGGCGGGGTGAGCAAGGGCGA"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     CDS             1925..2392
                     /label="GFP-N"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     CDS             1925..3221
                     /label="Translation 1925-3221"
     primer          complement(2047..2066)
                     /label="RG667"
                     /note="sequence: AGATGAACTTCAGGGTCAGC"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer          2292..2311
                     /label="RG617"
                     /note="sequence: TCGAGCTGAAGGGCATCGAC"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          2336..2355
                     /label="RG672"
                     /note="sequence: GGGCACAAGCTGGAGTACAA"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     primer          2353..2372
                     /label="RG673"
                     /note="sequence: CAACTACAACAGCCACAACG"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer          complement(2372..2412)
                     /label="AV469"
                     /note="sequence: GACAAAAAGACTAAACTTACCTTGTCGGCCATGATATAGAC"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     Splice Donor    2393..2394
                     /label="SD"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     Intron          2393..2524
                     /label="Intron loxP"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer          2393..2524
                     /label="AV467"
                     /note="sequence: GTAAGTTTAGTCTTTTTGTCTTTTATTTCAGGTCCCGATAACTTCGTATAATGTATGCTATACGAAGTTATGGATCCGGTGGTGGTGCAAATCAAAGAACTGCTCCTCAGTGGATGTTGCCTTTACTTCTAG"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          complement(2393..2524)
                     /label="AV468"
                     /note="sequence: CTAGAAGTAAAGGCAACATCCACTGAGGAGCAGTTCTTTGATTTGCACCACCACCGGATCCATAACTTCGTATAGCATACATTATACGAAGTTATCGGGACCTGAAATAAAAGACAAAAAGACTAAACTTAC"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     CDS             2393..2528
                     /label="Translation 2393-2528"
     primer          2423..2445
                     /label="AV533"
                     /note="sequence: GGTCCCGATAACTTCGTATAATG"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     loxP site       2430..2463
                     /label="loxP"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     primer          complement(2504..2524)
                     /label="AV555"
                     /note="sequence: CTAGAAGTAAAGGCAACATCC"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          complement(2504..2526)
                     /label="AV568"
                     /note="sequence: CACTAGAAGTAAAGGCAACATCC"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     Splice acceptor 2523..2524
                     /label="SA"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     primer          2527..2547
                     /label="AV554"
                     /note="sequence: AGGTGAGCAAGGGCGAGGCAG"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     CDS             2528..2528
                     /label="EGFP-C-PORTION"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     CDS             2529..3221
                     /label="mScarlet"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     CDS             2529..3221
                     /label="Translation 2529-3221"
     primer          2939..2961
                     /label="MZ497"
                     /note="sequence: GAAGAAGACAATGGGCTGGGAAG"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer          3102..3123
                     /label="AV556"
                     /note="sequence: GCCTACAACGTCGACCGCAAGT"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     primer          3199..3244
                     /label="AV534"
                     /note="sequence: GCGGCATGGACGAGCTGTACAAGTAGAACTTGTTTATTGCAGCTTA"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer          complement(3199..3244)
                     /label="AV535"
                     /note="sequence: TAAGCTGCAATAAACAAGTTCTACTTGTACAGCTCGTCCATGCCGC"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer          3222..3250
                     /label="AV487"
                     /note="sequence: TAGAACTTGTTTATTGCAGCTTATAATGG"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     polyA signal    3225..3355
                     /label="SV40-polyA"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     primer          3300..3319
                     /label="RG660"
                     /note="sequence: TCACTGCATTCTAGTTGTGG"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     primer_bind     complement(3372..3391)
                     /label="T3"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer_bind     complement(3517..3538)
                     /label="T7"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer_bind     complement(3537..3557)
                     /label="M13-rev"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     CDS             3670..4476
                     /label="KanR"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     gene            3670..4479
                     /label="KanR gene"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     CDS             3670..4479
                     /label="KanR CDS"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     rep_origin      complement(4599..5272)
                     /label="pUC origin of replication gene           711..2333 /gene=%22NEO_T2a_eGFP"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
ORIGIN
        1 CTTTCCTGCG TTATCCCCTG ATTCTGTGGA TAACCGTATT ACCGCCTTTG AGTGAGCTGA
       61 TACCGCTCGC CGCAGCCGAA CGACCGAGCG CAGCGAGTCA GTGAGCGAGG AAGCGGAAGA
      121 GCGCCCAATA CGCAAACCGC CTCTCCCCGC GCGTTGGCCG ATTCATTAAT GCAGCTGGCA
      181 CGACAGGTTT CCCGACTGGA AAGCGGGCAG TGAGCGCAAC GCAATTAATA CGCGTACCGC
      241 TAGCCAGGAA GAGTTTGTAG AAACGCAAAA AGGCCATCCG TCAGGATGGC CTTCTGCTTA
      301 GTTTGATGCC TGGCAGTTTA TGGCGGGCGT CCTGCCCGCC ACCCTCCGGG CCGTTGCTTC
      361 ACAACGTTCA AATCCGCTCC CGGCGGATTT GTCCTACTCA GGAGAGCGTT CACCGACAAA
      421 CAACAGATAA AACGAAAGGC CCAGTCTTCC GACTGAGCCT TTCGTTTTAT TTGATGCCTG
      481 GCAGTTCCCT ACTCTCGCGT TAACGCTAGC ATGGATGTTT TCCCAGTCAC GACGTTGTAA
      541 AACGACGGCC AGTGAATTGG attggctccg gtgcccgtca gtgggcagag cgcacatcgc
      601 ccacagtccc cgagaagttg gggggagggg tcggcaattg atccggtgcc tagagaaggt
      661 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
      721 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
      781 ccgccagaac acaggaccgg ttctagagcg ctcgcccgcc ccacgagagA TGaaaaagcc
      841 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga
      901 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
      961 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
     1021 tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaatttag
     1081 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     1141 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc
     1201 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     1261 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     1321 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     1381 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     1441 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     1501 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     1561 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
     1621 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     1681 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     1741 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
     1801 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aaGGCAGTGG
     1861 AGAGGGCAGA GGAAGTCTGC TAACATGCGG TGACGTCGAG GAGAATCCTG GCCCAGGGGG
     1921 CGGGGTGAGC AAGGGCGAGG AGCTGTTCAC CGGGGTGGTG CCCATCCTGG TCGAGCTGGA
     1981 CGGCGACGTA AACGGCCACA AGTTCAGCGT GTCCGGCGAG GGCGAGGGCG ATGCCACCTA
     2041 CGGCAAGCTG ACCCTGAAGT TCATCTGCAC CACCGGCAAG CTGCCCGTGC CCTGGCCCAC
     2101 CCTCGTGACC ACCCTGACCT ACGGCGTGCA GTGCTTCAGC CGCTACCCCG ACCACATGAA
     2161 GCAGCACGAC TTCTTCAAGT CCGCCATGCC CGAAGGCTAC GTCCAGGAGC GCACCATCTT
     2221 CTTCAAGGAC GACGGCAACT ACAAGACCCG CGCCGAGGTG AAGTTCGAGG GCGACACCCT
     2281 GGTGAACCGC ATCGAGCTGA AGGGCATCGA CTTCAAGGAG GACGGCAACA TCCTGGGGCA
     2341 CAAGCTGGAG TACAACTACA ACAGCCACAA CGTCTATATC ATGGCCGACA AGGTAAGTTT
     2401 AGTCTTTTTG TCTTTTATTT CAGGTCCCGA TAACTTCGTA TAATGTATGC TATACGAAGT
     2461 TATGGATCCG GTGGTGGTGC AAATCAAAGA ACTGCTCCTC AGTGGATGTT GCCTTTACTT
     2521 CTAGTGAGGT GAGCAAGGGC GAGGCAGTGA TCAAGGAGTT CATGCGGTTC AAGGTGCACA
     2581 TGGAGGGCTC CATGAACGGC CACGAGTTCG AGATCGAGGG CGAGGGCGAG GGCCGCCCCT
     2641 ACGAGGGCAC CCAGACCGCC AAGCTGAAGG TGACCAAGGG TGGCCCCCTG CCCTTCTCCT
     2701 GGGACATCCT GTCCCCTCAG TTCATGTACG GCTCCAGGGC CTTCACCAAG CACCCCGCCG
     2761 ACATCCCCGA CTACTATAAG CAGTCCTTCC CCGAGGGCTT CAAGTGGGAG CGCGTGATGA
     2821 ACTTCGAGGA CGGCGGCGCC GTGACCGTGA CCCAGGACAC CTCCCTGGAG GACGGCACCC
     2881 TGATCTACAA GGTGAAGCTC CGCGGCACCA ACTTCCCTCC TGACGGCCCC GTAATGCAGA
     2941 AGAAGACAAT GGGCTGGGAA GCGTCCACCG AGCGGTTGTA CCCCGAGGAC GGCGTGCTGA
     3001 AGGGCGACAT TAAGATGGCC CTGCGCCTGA AGGACGGCGG CCGCTACCTG GCGGACTTCA
     3061 AGACCACCTA CAAGGCCAAG AAGCCCGTGC AGATGCCCGG CGCCTACAAC GTCGACCGCA
     3121 AGTTGGACAT CACCTCCCAC AACGAGGACT ACACCGTGGT GGAACAGTAC GAACGCTCCG
     3181 AGGGCCGCCA CTCCACCGGC GGCATGGACG AGCTGTACAA GTAGAACTTG TTTATTGCAG
     3241 CTTATAATGG TTACAAATAA AGCAATAGCA TCACAAATTT CACAAATAAA GCATTTTTTT
     3301 CACTGCATTC TAGTTGTGGT TTGTCCAAAC TCATCAATGT ATCTTATCAT GTCTGGAGCT
     3361 CCAGCTTTTG TTCCCTTTAG TGAGGGTTAA TTTCGAGCTT ACCCAGCTTT CTTGTACAAA
     3421 GTTGGCATTA TAAGAAAGCA TTGCTTATCA ATTTGTTGCA ACGAACAGGT CACTATCAGT
     3481 CAAAATAAAA TCATTATTTG CCATCCAGCT GATATCCCCT ATAGTGAGTC GTATTACATG
     3541 GTCATAGCTG TTTCCTGGCA GCTCTGGCCC GTGTCTCAAA ATCTCTGATG TTACATTGCA
     3601 CAAGATAAAA ATATATCATC ATGAACAATA AAACTGTCTG CTTACATAAA CAGTAATACA
     3661 AGGGGTGTTA TGAGCCATAT TCAACGGGAA ACGTCGAGGC CGCGATTAAA TTCCAACATG
     3721 GATGCTGATT TATATGGGTA TAAATGGGCT CGCGATAATG TCGGGCAATC AGGTGCGACA
     3781 ATCTATCGCT TGTATGGGAA GCCCGATGCG CCAGAGTTGT TTCTGAAACA TGGCAAAGGT
     3841 AGCGTTGCCA ATGATGTTAC AGATGAGATG GTCAGACTAA ACTGGCTGAC GGAATTTATG
     3901 CCTCTTCCGA CCATCAAGCA TTTTATCCGT ACTCCTGATG ATGCATGGTT ACTCACCACT
     3961 GCGATCCCCG GAAAAACAGC ATTCCAGGTA TTAGAAGAAT ATCCTGATTC AGGTGAAAAT
     4021 ATTGTTGATG CGCTGGCAGT GTTCCTGCGC CGGTTGCATT CGATTCCTGT TTGTAATTGT
     4081 CCTTTTAACA GCGATCGCGT ATTTCGTCTC GCTCAGGCGC AATCACGAAT GAATAACGGT
     4141 TTGGTTGATG CGAGTGATTT TGATGACGAG CGTAATGGCT GGCCTGTTGA ACAAGTCTGG
     4201 AAAGAAATGC ATAAACTTTT GCCATTCTCA CCGGATTCAG TCGTCACTCA TGGTGATTTC
     4261 TCACTTGATA ACCTTATTTT TGACGAGGGG AAATTAATAG GTTGTATTGA TGTTGGACGA
     4321 GTCGGAATCG CAGACCGATA CCAGGATCTT GCCATCCTAT GGAACTGCCT CGGTGAGTTT
     4381 TCTCCTTCAT TACAGAAACG GCTTTTTCAA AAATATGGTA TTGATAATCC TGATATGAAT
     4441 AAATTGCAGT TTCATTTGAT GCTCGATGAG TTTTTCTAAT CAGAATTGGT TAATTGGTTG
     4501 TAACACTGGC AGAGCATTAC GCTGACTTGA CGGGACGGCG CAAGCTCATG ACCAAAATCC
     4561 CTTAACGTGA GTTACGCGTC GTTCCACTGA GCGTCAGACC CCGTAGAAAA GATCAAAGGA
     4621 TCTTCTTGAG ATCCTTTTTT TCTGCGCGTA ATCTGCTGCT TGCAAACAAA AAAACCACCG
     4681 CTACCAGCGG TGGTTTGTTT GCCGGATCAA GAGCTACCAA CTCTTTTTCC GAAGGTAACT
     4741 GGCTTCAGCA GAGCGCAGAT ACCAAATACT GTTCTTCTAG TGTAGCCGTA GTTAGGCCAC
     4801 CACTTCAAGA ACTCTGTAGC ACCGCCTACA TACCTCGCTC TGCTAATCCT GTTACCAGTG
     4861 GCTGCTGCCA GTGGCGATAA GTCGTGTCTT ACCGGGTTGG ACTCAAGACG ATAGTTACCG
     4921 GATAAGGCGC AGCGGTCGGG CTGAACGGGG GGTTCGTGCA CACAGCCCAG CTTGGAGCGA
     4981 ACGACCTACA CCGAACTGAG ATACCTACAG CGTGAGCTAT GAGAAAGCGC CACGCTTCCC
     5041 GAAGGGAGAA AGGCGGACAG GTATCCGGTA AGCGGCAGGG TCGGAACAGG AGAGCGCACG
     5101 AGGGAGCTTC CAGGGGGAAA CGCCTGGTAT CTTTATAGTC CTGTCGGGTT TCGCCACCTC
     5161 TGACTTGAGC GTCGATTTTT GTGATGCTCG TCAGGGGGGC GGAGCCTATG GAAAAACGCC
     5221 AGCAACGCGG CCTTTTTACG GTTCCTGGCC TTTTGCTGGC CTTTTGCTCA CATGTT
//