LOCUS       Addgene_submission:_L0- 4938 bp ds-DNA     circular     19-MAR-2024
DEFINITION  .
ACCESSION   urn.local...1531424985373.147
VERSION     urn.local...1531424985373.147
COMMENT     Imported using the Genbank importer. File name: pblunt.ape
            ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     misc_feature    1611..1631
                     /label=">pNL_oligo_2699"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    1611..1631
                     /label=">pNL_oligo_2699"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    1611..1634
                     /label=">pNL_oligo_2215"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    1611..1634
                     /label=">pNL_oligo_2215"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer_bind     complement(1611..1634)
                     /label=">pNL_oligo_2216"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     complement(1611..1634)
                     /label=">pNL_oligo_2216"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer_bind     complement(1613..1634)
                     /label=">pNL_oligo_3070"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer_bind     complement(1613..1634)
                     /label=">pNL_oligo_3070"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    1632..1654
                     /label=">pNL_oligo_552"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    1632..1654
                     /label=">pNL_oligo_552"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    1638..1653
                     /label=">pNL_oligo_37"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    1638..1653
                     /label=">pNL_oligo_37"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    1647..1672
                     /label=">pNL_oligo_1548"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    1647..1672
                     /label=">pNL_oligo_1548"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    1661..1679
                     /label=">pNL_oligo_3104"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    1661..1679
                     /label=">pNL_oligo_3104"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    1662..1678
                     /label=">pNL_oligo_16"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    1662..1678
                     /label=">pNL_oligo_16"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    1692..1713
                     /label=">pNL_oligo_1301"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    1692..1713
                     /label=">pNL_oligo_1301"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    1705..1719
                     /label=">pNL_oligo_2447"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    1705..1719
                     /label=">pNL_oligo_2447"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          1727..1770
                     /label="pBLUNT:rbcS_gibsonfwd"
                     /note="sequence: ggatatctgcagaattcaggcactctgtggtctcagcagcgatt"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer          complement(1727..1770)
                     /label="oNL5961_pBLUNT:rbcS_gibsonrev"
                     /note="sequence: aatcgctgctgagaccacagagtgcctgaattctgcagatatcc"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    1747..1761
                     /label="BsaI linker"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     source          1747..3172
                     /label="source"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    1762..1765
                     /label="GG3"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    complement(1766..2412)
                     /label="rbcS terminator_48839"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          2394..2453
                     /label="mCherryNLS:rbcS_gibsonfwd"
                     /note="sequence: tgatacgaacgaaagctttgactttcctcttcttctttggttaagatctgtacagctcgt"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer          complement(2394..2453)
                     /label="mCherryNLS:rbcS_gibsonrev"
                     /note="sequence: acgagctgtacagatcttaaccaaagaagaagaggaaagtcaaagctttcgttcgtatca"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer          2396..2455
                     /label="oNL5972_mCherryNLS:rbcS_gibsonfwd_new"
                     /note="sequence: atacgaacgaaagctttgactttcctcttcttctttggttaagatctgtacagctcgtcc"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    complement(2413..2433)
                     /label="SV40 NLS 3'"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     CDS             complement(2434..3147)
                     /label="mCherry"
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_fwdcolor="#ff0000"
     primer_bind     complement(2529..2554)
                     /label="sJAB007"
                     /ApEinfo_revcolor="#6666ff"
                     /ApEinfo_fwdcolor="#6666ff"
     primer          3128..3192
                     /label="oNL5958_mCherryNLS:pBLUNT_gibsonfwd"
                     /note="sequence: tcctcgcccttgctcaccattgtttagctttgagacctcatcacgcctgaattccagcacactgg"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          complement(3128..3192)
                     /label="mCherryNLS:pBLUNT_gibsonrev"
                     /note="sequence: ccagtgtgctggaattcaggcgtgatgaggtctcaaagctaaacaatggtgagcaagggcgagga"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          complement(3131..3190)
                     /label="oNL5978_mCherryNLS:pBLUNT_gibsonrev_new"
                     /note="sequence: agtgtgctggaattcaggcgtgatgaggtctcaaagctaaacaatggtgagcaagggcga"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    complement(3148..3153)
                     /label="Short enhancer for gene expression"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    3154..3157
                     /label="GG4"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    complement(3158..3172)
                     /label="BsaI linker4"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    3173..3505
                     /label="Digestion of pBlunt_BB (reversed)(concatenatedsequence 1)"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer_bind     complement(3281..3309)
                     /label=">pNL_oligo_553"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer_bind     complement(3281..3309)
                     /label=">pNL_oligo_553"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer_bind     complement(3285..3301)
                     /label=">pNL_oligo_38"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer_bind     complement(3285..3301)
                     /label=">pNL_oligo_38"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    3290..3318
                     /label=">pNL_oligo_2085"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    3290..3318
                     /label=">pNL_oligo_2085"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    3506..1746
                     /label="Digestion of pBlunt_BB (reversed)(concatenatedsequence 1)"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
ORIGIN
        1 gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa taattcagaa
       61 gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta
      121 aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc
      181 caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga
      241 aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag
      301 atcctcgccg tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc
      361 ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc
      421 tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg
      481 cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga
      541 caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac
      601 aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc
      661 ctcgtcttgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg
      721 cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca
      781 gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg
      841 ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca gagcttgatc ccctgcgcca
      901 tcagatcctt ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc
      961 agagggcgcc ccagctggca attccggttc gcttgctgtc cataaaaccg cccagtctag
     1021 ctatcgccat gtaagcccac tgcaagctac ctgctttctc tttgcgcttg cgttttccct
     1081 tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc
     1141 tttctacgtg aaaaggatct aggtgaagat cctttttgat aatctcatgc ctgacattta
     1201 tattccccag aacatcaggt taatggcgtt tttgatgtca ttttcgcggt ggctgagatc
     1261 agccacttct tccccgataa cggagaccgg cacactggcc atatcggtgg tcatcatgcg
     1321 ccagctttca tccccgatat gcaccaccgg gtaaagttca cgggagactt tatctgacag
     1381 cagacgtgca ctggccaggg ggatcaccat ccgtcgcccc ggcgtgtcaa taatatcact
     1441 ctgtacatcc acaaacagac gataacggct ctctctttta taggtgtaaa ccttaaactg
     1501 ccgtacgtat aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     1561 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     1621 ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact cactataggg
     1681 cgaattgggc cctctagatg catgctcgag cggccgccag tgtgatggat atctgcagaa
     1741 ttcaggcact ctgtggtctc agcagcgatt gatgcatgtt gtcaatcaat tggcaagtca
     1801 taaaatgcat taaaaaatat tttcatactc aactacaaat ccatgagtat aactataatt
     1861 ataaagcaat gattagaatc tgacaaggat tctggaaaat tacataaagg aaagttcata
     1921 aatgtctaaa acacaagagg acatacttgt attcagtaac atttgcagct tttctaggtc
     1981 tgaaaatata tttgttgcct agtgaataag cataatggta caactacaag tgttttactc
     2041 ctcatattaa cttcggtcat tagaggccac gatttgacac atttttactc aaaacaaaat
     2101 gtttgcatat ctcttataat ttcaaattca acacacaaca aataagagaa aaaacaaata
     2161 atattaattt gagaatgaac aaaaggacca tatcattcat taactcttct ccatccattt
     2221 ccatttcaca gttcgatagc gaaaaccgaa taaaaaacac agtaaattac aagcacaaca
     2281 aatggtacaa gaaaaacagt tttcccaatg ccataatact caaactcagt aggattctgg
     2341 tgtgtgcgca atgaaactga tgcattgaac ttgacgaacg ttgtcgaaac cgatgatacg
     2401 aacgaaagct ttgactttcc tcttcttctt tggttaagat ctgtacagct cgtccatgcc
     2461 gccggtggag tggcggccct cggcgcgttc gtactgttcc acgatggtgt agtcctcgtt
     2521 gtgggaggtg atgtccaact tgatgttgac gttgtaggcg ccgggcagct gcacgggctt
     2581 cttggccttg taggtggtct tgacctcagc gtcgtagtgg ccgccgtcct tcagcttcag
     2641 cctctgcttg atctcgccct tcagggcgcc gtcctcgggg tacatccgct cggaggaggc
     2701 ctcccagccc atggtcttct tctgcattac ggggccgtcg gaggggaagt tggtgccgcg
     2761 cagcttcacc ttgtagatga actcgccgtc ctgcagggag gagtcctggg tcacggtcac
     2821 cacgccgccg tcctcgaagt tcatcacgcg ctcccacttg aagccctcgg ggaaggacag
     2881 cttcaagtag tcggggatgt cggcggggtg cttcacgtag gccttggagc cgtacatgaa
     2941 ctgaggggac aggatgtccc aggcgaaggg cagggggcca cccttggtca ccttcagctt
     3001 ggcggtctgg gtgccctcgt aggggcggcc ctcgccctcg ccctcgatct cgaactcgtg
     3061 gccgttcacg gagccctcca tgtgcacctt gaagcgcatg aactccttga tgatggccat
     3121 gttatcctcc tcgcccttgc tcaccattgt ttagctttga gacctcatca cgcctgaatt
     3181 ccagcacact ggcggccgtt actagtggat ccgagctcgg taccaagctt gatgcatagc
     3241 ttgagtattc taacgcgtca cctaaatagc ttggcgtaat catggtcata gctgtttcct
     3301 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3361 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3421 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3481 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3541 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3601 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3661 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3721 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3781 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3841 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3901 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3961 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4021 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4081 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4141 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4201 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4261 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4321 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4381 cttttaaatt aaaaatgaag ttttagcacg tgtcagtcct gctcctcggc cacgaagtgc
     4441 acgcagttgc cggccgggtc gcgcagggcg aactcccgcc cccacggctg ctcgccgatc
     4501 tcggtcatgg ccggcccgga ggcgtcccgg aagttcgtgg acacgacctc cgaccactcg
     4561 gcgtacagct cgtccaggcc gcgcacccac acccaggcca gggtgttgtc cggcaccacc
     4621 tggtcctgga ccgcgctgat gaacagggtc acgtcgtccc ggaccacacc ggcgaagtcg
     4681 tcctccacga agtcccggga gaacccgagc cggtcggtcc agaactcgac cgctccggcg
     4741 acgtcgcgcg cggtgagcac cggaacggca ctggtcaact tggccatggt ggccctcctc
     4801 acgtgctatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4861 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4921 gatgcggtgt gaaatacc
//