LOCUS       Addgene_submission:_L0I 5053 bp ds-DNA     linear       19-MAR-2024
DEFINITION  .
ACCESSION   New Construct
KEYWORDS    "ID" "File Name" "Description" "GoldenGate" "Part/Tool"
            "Resistance" "Backbone"
SOURCE      Synthetic
  ORGANISM  Synthetic
COMMENT     Imported using the Genbank importer. File name:
            Level0_-_2023-01-05.gb Imported using the Genbank importer. File
            name: PhiC31-stop GG1-GG4 level0.gb
FEATURES             Location/Qualifiers
     misc_feature    1..15
                     /label="BsaI linker2"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     source          1..1541
                     /label="source"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    16..16
                     /label="GG1"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    17..1516
                     /label="Bxb1 without NLS"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             17..1516
                     /label="Translation 17-1516"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MRALVVIRLSRVTDATTSPERQLESCQQLCAQRGWDVVGVAEDLDVSGAVDPFDRKRRPNLARWLAFEEQPFDVIVAYRVDRLTRSIRHLQQLVHWAEDHKKLVVSATEAHFDTTTPFAAVVIALMGTVAQMELEAIKERNRSAAHFNIRAGKYRGSLPPWGYLPTRVDGEWRLVPDPVQRERILEVYHRVVDNHEPLHLVAHDLNRRGVLSPKDYFAQLQGREPQGREWSATALKRSMISEAMLGYATLNGKTVRDDDGAPLVRAEPILTREQLEALRAELVKTSRAKPAVSTPSLLLRVLFCAVCGEPAYKFAGGGRKHPRYRCRSMGFPKHCGNGTVAMAEWDAFCEEQVLDLLGDAERLEKVWVAGSDSAVELAEVNAELVDLTSLIGSPAYRAGSPQREALDARIAALAARQEELEGLEARPSGWEWRETGQRFGDWWREQDTAAKNTWLRSMNVRLTFDVRGGLTRTIDFGDLQEYEQHLRLGSVVERLHTGMS"
     primer          complement(1290..1307)
                     /label="oNL6103_Bxb1_mutagenesis_R"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="sequence: CCCAGCCAGACGGGCGAG"
     primer          complement(1290..1307)
                     /label="oNL6103_Bxb1_mutagenesis_R"
                     /note="sequence: CCCAGCCAGACGGGCGAG"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          1308..1328
                     /label="oNL6102_Bxb1_mutagenesis_F"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
                     /note="sequence: AGTGGCGCGAaACCGGGCAGC"
     primer          1308..1328
                     /label="oNL6102_Bxb1_mutagenesis_F"
                     /note="sequence: AGTGGCGCGAaACCGGGCAGC"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    1517..1522
                     /label="Stop codon"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    1523..1526
                     /label="GG4"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    complement(1527..1541)
                     /label="BsaI linker4"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    complement(1542..4720)
                     /label="Digestion of pBlunt_BB (reversed)(concatenatedsequence 1)"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    complement(1569..1583)
                     /label=">pNL_oligo_2447"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    complement(1569..1583)
                     /label=">pNL_oligo_2447"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    complement(1575..1596)
                     /label=">pNL_oligo_1301"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    complement(1575..1596)
                     /label=">pNL_oligo_1301"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    complement(1609..1627)
                     /label=">pNL_oligo_3104"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    complement(1609..1627)
                     /label=">pNL_oligo_3104"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    complement(1610..1626)
                     /label=">pNL_oligo_16"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    complement(1610..1626)
                     /label=">pNL_oligo_16"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    complement(1616..1641)
                     /label=">pNL_oligo_1548"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    complement(1616..1641)
                     /label=">pNL_oligo_1548"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    complement(1634..1656)
                     /label=">pNL_oligo_552"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    complement(1634..1656)
                     /label=">pNL_oligo_552"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    complement(1635..1650)
                     /label=">pNL_oligo_37"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    complement(1635..1650)
                     /label=">pNL_oligo_37"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer_bind     1654..1675
                     /label=">pNL_oligo_3070"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer_bind     1654..1675
                     /label=">pNL_oligo_3070"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer_bind     1654..1677
                     /label=">pNL_oligo_2216"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer_bind     1654..1677
                     /label=">pNL_oligo_2216"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    complement(1654..1677)
                     /label=">pNL_oligo_2215"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    complement(1654..1677)
                     /label=">pNL_oligo_2215"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    complement(1657..1677)
                     /label=">pNL_oligo_2699"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    complement(1657..1677)
                     /label=">pNL_oligo_2699"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    complement(4721..5053)
                     /label="Digestion of pBlunt_BB (reversed)(concatenatedsequence 1)"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    complement(4908..4936)
                     /label=">pNL_oligo_2085"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    complement(4908..4936)
                     /label=">pNL_oligo_2085"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer_bind     4917..4945
                     /label=">pNL_oligo_553"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer_bind     4917..4945
                     /label=">pNL_oligo_553"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer_bind     4925..4941
                     /label=">pNL_oligo_38"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer_bind     4925..4941
                     /label=">pNL_oligo_38"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
ORIGIN
        1 ggatcggagg tctcaaatga gagccctggt agtcatccgc ctgtcccgcg tcaccgatgc
       61 tacgacttca ccggagcgtc agctggagtc ttgccagcag ctctgcgccc agcgcggctg
      121 ggacgtcgtc ggggtagcgg aggatctgga cgtctccggg gcggtcgatc cgttcgaccg
      181 gaagcgcaga ccgaacctgg cccggtggct tgcgttcgag gagcaaccgt ttgacgtgat
      241 cgtggcgtac cgggtagatc ggttgacccg atcgatccgg catcttcagc agctggtcca
      301 ctgggccgag gaccacaaga agctggtcgt ctccgcgacc gaagcgcact tcgatacgac
      361 gacgccgttt gcggcggtcg tcatcgcgct tatgggaacg gtggcgcaga tggaattaga
      421 agcgatcaaa gagcggaacc gttcggctgc gcatttcaat atccgcgccg ggaaataccg
      481 agggtccctg ccgccgtggg gatacctgcc tacgcgcgtg gacggggagt ggcgactggt
      541 gccggaccct gtgcagcgag agcgcatcct cgaggtgtat caccgcgtcg tcgacaacca
      601 cgagccgctg catctggtgg cccacgacct gaaccggcgt ggtgtcctgt cgccgaagga
      661 ctacttcgcg cagctgcaag gccgcgagcc gcagggccgg gagtggtcgg ctaccgcgct
      721 gaagcgatcg atgatctccg aggcgatgct cgggtacgcg actctgaacg gtaagaccgt
      781 ccgagacgac gacggagccc cgctggtgcg ggctgagccg atcctgaccc gtgagcagct
      841 ggaggcgctg cgcgccgagc tcgtgaagac ctcccgggcg aagcccgcgg tgtctacccc
      901 gtcgctgctg ctgcgggtgt tgttctgcgc ggtgtgcggg gagcccgcgt acaagttcgc
      961 cgggggagga cgtaagcacc cgcgctaccg ctgccgctcg atggggttcc cgaagcactg
     1021 cgggaacggc acggtggcga tggccgagtg ggacgcgttc tgcgaggagc aggtactgga
     1081 tctgctcggg gacgcggagc gtctggagaa agtctgggta gcgggctcgg actccgcggt
     1141 cgaactcgcg gaggtgaacg cggagctggt ggacctgacg tcgctgatcg gctccccggc
     1201 ctaccgggcg ggctctccgc agcgagaagc actggatgcc cgtattgcgg cgctggccgc
     1261 gcggcaagag gagctggagg gcctggaggc tcgcccgtct ggctgggagt ggcgcgaaac
     1321 cgggcagcgg ttcggggact ggtggcggga gcaggacacc gcggcaaaga acacctggct
     1381 tcggtcgatg aacgttcggc tgacgttcga cgtccgcggc gggctgactc gcacgatcga
     1441 cttcggggat cttcaggagt acgagcagca tctcaggctc ggcagcgtgg tcgaacggct
     1501 acacaccggg atgtcgtaat aagctttgag acctcatcac gcctgaattc tgcagatatc
     1561 catcacactg gcggccgctc gagcatgcat ctagagggcc caattcgccc tatagtgagt
     1621 cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     1681 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     1741 cccgcaccga tcgcccttcc caacagttgc gcagcctata cgtacggcag tttaaggttt
     1801 acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg
     1861 acacgccggg gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag
     1921 tctcccgtga actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca
     1981 ccgatatggc cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc
     2041 gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggca
     2101 tgagattatc aaaaaggatc ttcacctaga tccttttcac gtagaaagcc agtccgcaga
     2161 aacggtgctg accccggatg aatgtcagct actgggctat ctggacaagg gaaaacgcaa
     2221 gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg
     2281 ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga
     2341 agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat
     2401 caagctctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     2461 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     2521 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     2581 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     2641 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     2701 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     2761 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     2821 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     2881 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     2941 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     3001 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3061 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3121 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3181 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgaattatta
     3241 acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3301 gcatcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3361 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatag
     3421 cacgtgagga gggccaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc
     3481 gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg
     3541 gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag
     3601 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg
     3661 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg
     3721 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac
     3781 tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc taaaacttca tttttaattt
     3841 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3901 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3961 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4021 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4081 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4141 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4201 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4261 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4321 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4381 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4441 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4501 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4561 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4621 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4681 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     4741 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     4801 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     4861 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     4921 cacacaggaa acagctatga ccatgattac gccaagctat ttaggtgacg cgttagaata
     4981 ctcaagctat gcatcaagct tggtaccgag ctcggatcca ctagtaacgg ccgccagtgt
     5041 gctggaattc agg
//