LOCUS       pSH1_PtxD_BHM2675       6361 bp ds-DNA     circular     30-MAY-2025
DEFINITION  .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
COMMENT     Imported using the Genbank importer. File name:
            addgene-plasmid-67940-sequence-120525 (1).gbk Complementation in
            Cryptococcus neoformans. Targets DNA constructs to a Safe Haven
            Site, a small gene-free region. Contains nourseothricin resistance
            marker (NAT).
FEATURES             Location/Qualifiers
     misc_feature    complement(13..762)
                     /label="Upstream homology"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    complement(791..1540)
                     /label="Downstream Homology"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    complement(1561..1787)
                     /label="TRP1 Terminator"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    1603..1624
                     /label="Adaptor for KO R"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_feature    complement(1635..1653)
                     /label="MJB556"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    complement(1815..2877)
                     /label="PtxD_CnOpt_Intron"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    complement(2487..2538)
                     /label="Intron"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    complement(2896..3638)
                     /label="ACT1 Promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    3646..3664
                     /label="MJB555"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    complement(3651..3668)
                     /label="Adaptor for KO F"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     CDS             complement(3851..4216)
                     /label="lacZ-alpha"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer_bind     3985..4007
                     /label="M13/pUC Forward"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer_bind     3999..4016
                     /label="M13 Forward"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     4000..4016
                     /label="M13 fwd"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     promoter        4026..4044
                     /label="T7 promoter"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer_bind     4026..4045
                     /label="T7"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    4053..4160
                     /label="MCS"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer_bind     4069..4085
                     /label="pBluescriptKS"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     4070..4086
                     /label="KS primer"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer_bind     complement(4120..4136)
                     /label="pBluescriptSK"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer_bind     complement(4120..4136)
                     /label="SK primer"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     promoter        complement(4173..4191)
                     /label="T3 promoter"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer_bind     complement(4173..4193)
                     /label="T3"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer_bind     complement(4212..4228)
                     /label="M13 Reverse"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer_bind     complement(4212..4228)
                     /label="M13 rev"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer_bind     complement(4225..4247)
                     /label="M13/pUC Reverse"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     protein_bind    4236..4252
                     /label="lac operator"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     promoter        complement(4260..4290)
                     /label="lac promoter"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     protein_bind    4305..4326
                     /label="CAP binding site"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer_bind     complement(4443..4460)
                     /label="L4440"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     rep_origin      complement(4614..5202)
                     /label="ori"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer_bind     complement(4694..4713)
                     /label="pBR322ori-F"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     CDS             complement(5373..6233)
                     /label="AmpR"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     5996..6015
                     /label="Amp-R"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     promoter        complement(6234..6338)
                     /label="AmpR promoter"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
ORIGIN
        1 ctcatcagat ctgcccatag acttcaaatc atggaattgg acgtttatct tcttgctttg
       61 gatatccacc ctatcgtaca atcttggcac tttggatcat acgaggcact ttgcggacta
      121 gggaatggtt atcatcaatt tcttggacct ctccatgaca cctggcatcg tcgagctacc
      181 acagccccct gaattcttaa aggcgaaccc cccttcctag gaaccttgag cgctggactg
      241 gatggcactc tgagaccgcc tatgcttaag gacgcgagct tacccttcca ttgatggaag
      301 tcgatcattt atgataatag aacacggcgg aacttgactt tacaggccaa gggaagcaac
      361 aggagaagaa gctaaggtta taacaggctc agcatatttg tatatacggt gccattcaac
      421 gctgaacact ttgtactcac ggtctgcccg ttgaataaca ccatctgctg atgatcatcc
      481 caatttgtga gtcttatttg ccattgatcc tggtgtataa ccgatactga agccctttag
      541 gtctgatcag ttatagacgt ggtatattca ttcttcctag attagctatg ccgaaagaga
      601 tggatacctt gatgtgactg gaatgccatg aagcaggaga ttgtagcagc tcaaattgga
      661 gaaggatgtt gcaaccgtcg gccgtgaagg tcttgtcata tctttgtctg cctgggagaa
      721 tatgtacaag tgccccgatg ttactgatgg aaaacatcgt gtggcgcgcc acgatacttg
      781 tgttaattaa accaaaccag tctgtgccaa gtggattgct tatgcatcga gtggcgagcc
      841 cactgacctc tgggcccctt ttttgaacta taaaaagcgg ccaagtctga tttcggtctc
      901 acaatcatca acatcgactt ctcttttcgc ctcgatcaac ctaaacaacg gtcactcagc
      961 catgaactgg acaatggaat atttcgcctt gccttggatc tttggcctct agactcaatc
     1021 aacgtcagga taaaggtgag cataggttgc tcactcactg tagccttttt ttcccttctt
     1081 cttggtccag atggttccca aggctctata aagtgtatct gtcagttcca cttccttact
     1141 ccgaagcgat tcagagttgt aaagtgcgtc tgaccccctg attatatctc aaagtccctt
     1201 cacaaattca gacatgatgc gataaagtgc cggattgtca ttcgcgtctt gtgactgtat
     1261 agaaagtgcc ggcatcctct tttcttttcc tctttggccg agcgactcag agttgtgaag
     1321 cctgctggta tctgtctgtt cccttgccct gataaagaat atcgaagagc atttctttga
     1381 ttcacaatac gttgacgaat ggtgtgatgg atcctaagca gcgccagccc tttgaacttg
     1441 tgtcgaaaaa aaaaaaaaaa cagcatatga tacgtaaagg gaagaaaagg atggagctga
     1501 ctgttttgac aggcataatt ttgagcgtgc ttcattggcc ggtttatctg tattaacacg
     1561 gaagagatgt agaaactagc ttcctggttt cagagacaag gagccatgaa gatcctgagg
     1621 attcaacggc gttgctgagg gtgggaagga tggtattgtt gtctgtagat agattaatgg
     1681 gcatataatg tcaaaaaaga tgattgacac gatgtctgtc tcgaattcgc agtgtgctaa
     1741 aattgtatca cttctctaat atgtatatat acaccctcta aggaaaatta accccttacc
     1801 gccttcacga attcttaaca ggcggcgggc tcggccttgg ggagtcggtt ggcggcgttg
     1861 atgggtcggg caccggcgag gacctggatg atgttctggg cggcacatcg ctcgatctcg
     1921 agtcggacgg ctcggacggc ggaaccgatg tggggggtga agagggtgtt ggggtgggcg
     1981 aggagggcgg ggtcgatgag tcggggtcgg tcggctcggg cccagtcctc catctcgaag
     2041 acgtcggcgg cgtaaccacc gagctgacct cgctcgaggg cggcgaggac ggcggcctcg
     2101 tcgacgacgg aacctcgaca ggggttgacg aggagggcac cgggtcggac gagggcgagg
     2161 agctcggcgt tgacgaggtg ctgggtgtcg gcgttgaggg ggagggcgag gaggatgaag
     2221 tcggaggagg cgaagagctc ggaacaggcg acctgtcgga gaccgagtcg ctgctcggtc
     2281 tgggtgtcga gggccttggc ctcgtggtac tggagggtgg caccccaacc ctggagtcgg
     2341 tcggccatgg caagacctat ggcacccata ccgaggatac cgacggtggc gttgtcgaga
     2401 ccggtaccgt agaactgggg ctgccaaccc tggaactcac cggatcggac gaaggcgtcg
     2461 gcggctcgga ggtgtcgacc gagaccctgt cgggagatgt tagtggagta acatgttgcg
     2521 ggaaacgggg ttacatacga cggcgagccc gatagcgagc tcggcggtgg ggacggtgag
     2581 gaggtcgggg acgaaggtga gccagacacc tcgggcggtg caggcatcga cgtcgaagtt
     2641 gtcgaaaccc ttgagggcac aaccgacgac tcggagctcg ggacaggcct ggaggaagtc
     2701 ggcgtcgact cggtcgggca tgaaggccat catggcctgg gcgtctcgac atcgtcggag
     2761 gatctcctct cgggtgaggg tggagtcggt ctggttggtc atgagctcac agtggggggc
     2821 gaggagctgg aggatctcgt cgtggactcg gtgggtgatg acgagcttgg ggagcatggc
     2881 ggccgccata gacatgttgg gcgagtttac taatggaaaa agacaattag caaagaggct
     2941 gcatattagg agacgcggtg aagattgggg aggacaatag ggtccgtgca ccaaaaagtc
     3001 ctgacggcag ctctgttcaa cgcacatcca tctcttccag ctatgcgtgt ggcaccagca
     3061 cggatggatt gttgcaccaa aaggacaagg cgagatggat gcaggtcgag gcgatgcggt
     3121 gctatcggcc ctcgttgtcg aacgccacgt cgtcctctcg gcgaccgtca atcctggaag
     3181 gaaatgacca agcgtgcacc cagcatccag catccaacat ccagcaccca gcacctagca
     3241 cccatgatcc aatggccggc atgcagatca gcgtggggag agcaccatct agcagcccct
     3301 ggcaggccca gcattgtgct cggatgtgcg ataggcggtg ggtgcgataa ggtggattag
     3361 tggaggatgg aggactgagt ggcgggcaag ggaagggggc gggaagatga ctcaccttgg
     3421 gcagtgggtg ggggatgcgg tataggggat gtatgtggat gctggcggag gatagaagct
     3481 gtaagctgag ctggtgccaa cggtgacgct gtgagagtgg tctcggcgac cctgacggcg
     3541 actcggcgtt accccgtgtg gcacaccgga cgcgcgtcga gtgacgttgc tgctattgtt
     3601 gttaccatca tcctctcctc cgacaaccat acactcagtg cacaccatgc cactcgaatc
     3661 ctgcatgctt atgtgagtcc tccccttccc gtgcgccgct ctccagctca catcctcgca
     3721 gccatggact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3781 cggcctattg gttaaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     3841 atattaacgc ttacaatttc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     3901 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     3961 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc
     4021 gcgcgtaata cgactcacta tagggcgaat tgggtaccgg gccccccctc gaggtcgacg
     4081 gtatcgataa gcttgatatc gaattcctgc agcccggggg atccactagt tctagagcgg
     4141 ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     4201 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4261 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4321 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4381 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4441 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4501 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4561 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4621 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4681 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4741 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4801 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4861 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4921 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4981 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5041 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5101 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5161 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5221 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5281 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5341 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5401 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5461 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5521 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5581 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5641 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5701 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5761 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5821 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5881 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5941 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6001 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6061 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6121 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6181 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6241 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6301 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6361 c
//