LOCUS       pSH1_BLE_BHM2672        5725 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..2241)
                     /label="BLE_CnOpt_Intron"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    complement(2064..2115)
                     /label="Intron"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    complement(2260..3002)
                     /label="ACT1 Promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    3010..3028
                     /label="MJB555"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    complement(3015..3032)
                     /label="Adaptor for KO F"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     CDS             complement(3215..3580)
                     /label="lacZ-alpha"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer_bind     3349..3371
                     /label="M13/pUC Forward"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer_bind     3363..3380
                     /label="M13 Forward"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     3364..3380
                     /label="M13 fwd"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     promoter        3390..3408
                     /label="T7 promoter"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer_bind     3390..3409
                     /label="T7"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    3417..3524
                     /label="MCS"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer_bind     3433..3449
                     /label="pBluescriptKS"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     3434..3450
                     /label="KS primer"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer_bind     complement(3484..3500)
                     /label="pBluescriptSK"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer_bind     complement(3484..3500)
                     /label="SK primer"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     promoter        complement(3537..3555)
                     /label="T3 promoter"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer_bind     complement(3537..3557)
                     /label="T3"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer_bind     complement(3576..3592)
                     /label="M13 Reverse"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer_bind     complement(3576..3592)
                     /label="M13 rev"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer_bind     complement(3589..3611)
                     /label="M13/pUC Reverse"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     protein_bind    3600..3616
                     /label="lac operator"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     promoter        complement(3624..3654)
                     /label="lac promoter"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     protein_bind    3669..3690
                     /label="CAP binding site"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer_bind     complement(3807..3824)
                     /label="L4440"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     rep_origin      complement(3978..4566)
                     /label="ori"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer_bind     complement(4058..4077)
                     /label="pBR322ori-F"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     CDS             complement(4737..5597)
                     /label="AmpR"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     5360..5379
                     /label="Amp-R"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     promoter        complement(5598..5702)
                     /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 attcttagtc ctgctcctcg gcgacgaagt ggacacagtt accggcgggg
     1861 tctcggaggg cgaactctcg accccagggc tgctcaccga tctcggtcat ggcgggaccg
     1921 gaggcgtctc ggaagttggt ggagacgacc tcggaccact cggcgtagag ctcgtcgaga
     1981 cctcggaccc agacccaggc gagggtgttg tcggggacga cctggtcctg gacggcggag
     2041 atgaagaggg tgacgtcgtc tcgctgtcgg gagatgttag tggagtaaca tgttgcggga
     2101 aacggggtta catacgacga caccggcgaa gtcgtcctcg acgaagtctc gggagaaacc
     2161 gagtcggtcg gtccagaact cgacggcacc ggcgacgtct cgggcggtga ggacggggac
     2221 ggcggaggtg agcttggcca tggcggccgc catagacatg ttgggcgagt ttactaatgg
     2281 aaaaagacaa ttagcaaaga ggctgcatat taggagacgc ggtgaagatt ggggaggaca
     2341 atagggtccg tgcaccaaaa agtcctgacg gcagctctgt tcaacgcaca tccatctctt
     2401 ccagctatgc gtgtggcacc agcacggatg gattgttgca ccaaaaggac aaggcgagat
     2461 ggatgcaggt cgaggcgatg cggtgctatc ggccctcgtt gtcgaacgcc acgtcgtcct
     2521 ctcggcgacc gtcaatcctg gaaggaaatg accaagcgtg cacccagcat ccagcatcca
     2581 acatccagca cccagcacct agcacccatg atccaatggc cggcatgcag atcagcgtgg
     2641 ggagagcacc atctagcagc ccctggcagg cccagcattg tgctcggatg tgcgataggc
     2701 ggtgggtgcg ataaggtgga ttagtggagg atggaggact gagtggcggg caagggaagg
     2761 gggcgggaag atgactcacc ttgggcagtg ggtgggggat gcggtatagg ggatgtatgt
     2821 ggatgctggc ggaggataga agctgtaagc tgagctggtg ccaacggtga cgctgtgaga
     2881 gtggtctcgg cgaccctgac ggcgactcgg cgttaccccg tgtggcacac cggacgcgcg
     2941 tcgagtgacg ttgctgctat tgttgttacc atcatcctct cctccgacaa ccatacactc
     3001 agtgcacacc atgccactcg aatcctgcat gcttatgtga gtcctcccct tcccgtgcgc
     3061 cgctctccag ctcacatcct cgcagccatg gactcaaccc tatctcggtc tattcttttg
     3121 atttataagg gattttgccg atttcggcct attggttaaa aaaatgagct gatttaacaa
     3181 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttccattcg ccattcaggc
     3241 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     3301 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     3361 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta
     3421 ccgggccccc cctcgaggtc gacggtatcg ataagcttga tatcgaattc ctgcagcccg
     3481 ggggatccac tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt
     3541 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3601 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     3661 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3721 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3781 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3841 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3901 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3961 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4021 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4081 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4141 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4201 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4261 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4321 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4381 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     4441 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4501 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4561 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4621 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4681 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4741 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4801 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4861 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4921 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4981 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5041 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5101 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5161 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5221 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5281 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5341 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5401 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5461 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5521 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5581 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5641 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5701 cgcacatttc cccgaaaagt gccac
//