LOCUS       pSH1_BSR_BHM2674        5773 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..2289)
                     /label="BSR_CnOpt_Intron"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    complement(2097..2148)
                     /label="Intron"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    complement(2308..3050)
                     /label="ACT1 Promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    3058..3076
                     /label="MJB555"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    complement(3063..3080)
                     /label="Adaptor for KO F"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     CDS             complement(3263..3628)
                     /label="lacZ-alpha"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer_bind     3397..3419
                     /label="M13/pUC Forward"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer_bind     3411..3428
                     /label="M13 Forward"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     3412..3428
                     /label="M13 fwd"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     promoter        3438..3456
                     /label="T7 promoter"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer_bind     3438..3457
                     /label="T7"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    3465..3572
                     /label="MCS"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer_bind     3481..3497
                     /label="pBluescriptKS"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     3482..3498
                     /label="KS primer"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer_bind     complement(3532..3548)
                     /label="SK primer"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer_bind     complement(3532..3548)
                     /label="pBluescriptSK"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     promoter        complement(3585..3603)
                     /label="T3 promoter"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer_bind     complement(3585..3605)
                     /label="T3"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer_bind     complement(3624..3640)
                     /label="M13 rev"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer_bind     complement(3624..3640)
                     /label="M13 Reverse"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer_bind     complement(3637..3659)
                     /label="M13/pUC Reverse"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     protein_bind    3648..3664
                     /label="lac operator"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     promoter        complement(3672..3702)
                     /label="lac promoter"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     protein_bind    3717..3738
                     /label="CAP binding site"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer_bind     complement(3855..3872)
                     /label="L4440"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     rep_origin      complement(4026..4614)
                     /label="ori"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer_bind     complement(4106..4125)
                     /label="pBR322ori-F"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     CDS             complement(4785..5645)
                     /label="AmpR"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     5408..5427
                     /label="Amp-R"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     promoter        complement(5646..5750)
                     /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 attcttagtt tcgggtgtac ttgaggggga tgagctcctc gatggtggtc
     1861 ttgacgagct taccgttcat ctcgatgagg acgaaacagt cgggggcgta gtcggagatg
     1921 agctctcgac acataccaca gggggagacg actcggatgg atcggtcgac ctcgtcggag
     1981 taggggtgtc ggacggcgac gatggtgtcg aagtccttct gaccgttgga gacggcggaa
     2041 ccgatggcga tggcctcggc acagacggtg actcgaccga tgtaggcctc gatgtgctgt
     2101 cgggagatgt tagtggagta acatgttgcg ggaaacgggg ttacatacga cggcggagat
     2161 gatctcaccg gtcttggttc ggatggcggc accgacgtgg tgcttgttgt cctcgtagag
     2221 catggtgatc ttctcggtgg cgacctcgac gagctcgagg tcctgctggg agatgttgaa
     2281 ggtcttcatg gcggccgcca tagacatgtt gggcgagttt actaatggaa aaagacaatt
     2341 agcaaagagg ctgcatatta ggagacgcgg tgaagattgg ggaggacaat agggtccgtg
     2401 caccaaaaag tcctgacggc agctctgttc aacgcacatc catctcttcc agctatgcgt
     2461 gtggcaccag cacggatgga ttgttgcacc aaaaggacaa ggcgagatgg atgcaggtcg
     2521 aggcgatgcg gtgctatcgg ccctcgttgt cgaacgccac gtcgtcctct cggcgaccgt
     2581 caatcctgga aggaaatgac caagcgtgca cccagcatcc agcatccaac atccagcacc
     2641 cagcacctag cacccatgat ccaatggccg gcatgcagat cagcgtgggg agagcaccat
     2701 ctagcagccc ctggcaggcc cagcattgtg ctcggatgtg cgataggcgg tgggtgcgat
     2761 aaggtggatt agtggaggat ggaggactga gtggcgggca agggaagggg gcgggaagat
     2821 gactcacctt gggcagtggg tgggggatgc ggtatagggg atgtatgtgg atgctggcgg
     2881 aggatagaag ctgtaagctg agctggtgcc aacggtgacg ctgtgagagt ggtctcggcg
     2941 accctgacgg cgactcggcg ttaccccgtg tggcacaccg gacgcgcgtc gagtgacgtt
     3001 gctgctattg ttgttaccat catcctctcc tccgacaacc atacactcag tgcacaccat
     3061 gccactcgaa tcctgcatgc ttatgtgagt cctccccttc ccgtgcgccg ctctccagct
     3121 cacatcctcg cagccatgga ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3181 ttttgccgat ttcggcctat tggttaaaaa aatgagctga tttaacaaaa atttaacgcg
     3241 aattttaaca aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt
     3301 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     3361 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     3421 cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc gggccccccc
     3481 tcgaggtcga cggtatcgat aagcttgata tcgaattcct gcagcccggg ggatccacta
     3541 gttctagagc ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta
     3601 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     3661 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     3721 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3781 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3841 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3901 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3961 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4021 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4081 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4141 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4201 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4261 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4321 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4381 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4441 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     4501 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4561 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4621 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4681 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4741 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4801 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4861 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4921 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4981 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5041 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5101 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5161 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5221 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5281 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5341 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5401 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5461 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5521 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5581 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5641 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5701 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5761 cgaaaagtgc cac
//