LOCUS       pSH1_BSD_BHM2673        5749 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..2265)
                     /label="BSD_CnOpt_Intron"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    complement(2082..2133)
                     /label="Intron"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    complement(2284..3026)
                     /label="ACT1 Promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    3034..3052
                     /label="MJB555"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    complement(3039..3056)
                     /label="Adaptor for KO F"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     CDS             complement(3239..3604)
                     /label="lacZ-alpha"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer_bind     3373..3395
                     /label="M13/pUC Forward"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer_bind     3387..3404
                     /label="M13 Forward"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     3388..3404
                     /label="M13 fwd"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     promoter        3414..3432
                     /label="T7 promoter"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer_bind     3414..3433
                     /label="T7"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    3441..3548
                     /label="MCS"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer_bind     3457..3473
                     /label="pBluescriptKS"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     3458..3474
                     /label="KS primer"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer_bind     complement(3508..3524)
                     /label="SK primer"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer_bind     complement(3508..3524)
                     /label="pBluescriptSK"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     promoter        complement(3561..3579)
                     /label="T3 promoter"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer_bind     complement(3561..3581)
                     /label="T3"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer_bind     complement(3600..3616)
                     /label="M13 rev"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer_bind     complement(3600..3616)
                     /label="M13 Reverse"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer_bind     complement(3613..3635)
                     /label="M13/pUC Reverse"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     protein_bind    3624..3640
                     /label="lac operator"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     promoter        complement(3648..3678)
                     /label="lac promoter"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     protein_bind    3693..3714
                     /label="CAP binding site"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer_bind     complement(3831..3848)
                     /label="L4440"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     rep_origin      complement(4002..4590)
                     /label="ori"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer_bind     complement(4082..4101)
                     /label="pBR322ori-F"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     CDS             complement(4761..5621)
                     /label="AmpR"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer_bind     5384..5403
                     /label="Amp-R"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     promoter        complement(5622..5726)
                     /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 attcttaacc ctcccagacg taaccggagg ggaggagctc tcggataccg
     1861 acggcggtgg gctgaccgtc ggagtccttg acgatggcct tgataccggg gtggaggtcg
     1921 aggaggacct gtcgacatcg accacagggg gagaggatac ctcggttctc gttaccgatg
     1981 gcgacgatac aggtgaggtt accggcggcg gcggcggcgg cggtaccgag gacgacgagc
     2041 tcggcacagg gaccaccggt gaagtggtag acgttgacac cctgtcggga gatgttagtg
     2101 gagtaacatg ttgcgggaaa cggggttaca tacggtgaag attcgaccgt cggaggagag
     2161 ggcggcggag gcgacggagt agtcctcgga gatggggatg gagttgatgg tggcggtggc
     2221 tcgctcgatg agggtggact cctcctggga gaggggcttg gccatggcgg ccgccataga
     2281 catgttgggc gagtttacta atggaaaaag acaattagca aagaggctgc atattaggag
     2341 acgcggtgaa gattggggag gacaataggg tccgtgcacc aaaaagtcct gacggcagct
     2401 ctgttcaacg cacatccatc tcttccagct atgcgtgtgg caccagcacg gatggattgt
     2461 tgcaccaaaa ggacaaggcg agatggatgc aggtcgaggc gatgcggtgc tatcggccct
     2521 cgttgtcgaa cgccacgtcg tcctctcggc gaccgtcaat cctggaagga aatgaccaag
     2581 cgtgcaccca gcatccagca tccaacatcc agcacccagc acctagcacc catgatccaa
     2641 tggccggcat gcagatcagc gtggggagag caccatctag cagcccctgg caggcccagc
     2701 attgtgctcg gatgtgcgat aggcggtggg tgcgataagg tggattagtg gaggatggag
     2761 gactgagtgg cgggcaaggg aagggggcgg gaagatgact caccttgggc agtgggtggg
     2821 ggatgcggta taggggatgt atgtggatgc tggcggagga tagaagctgt aagctgagct
     2881 ggtgccaacg gtgacgctgt gagagtggtc tcggcgaccc tgacggcgac tcggcgttac
     2941 cccgtgtggc acaccggacg cgcgtcgagt gacgttgctg ctattgttgt taccatcatc
     3001 ctctcctccg acaaccatac actcagtgca caccatgcca ctcgaatcct gcatgcttat
     3061 gtgagtcctc cccttcccgt gcgccgctct ccagctcaca tcctcgcagc catggactca
     3121 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3181 taaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     3241 acaatttcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     3301 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     3361 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg
     3421 actcactata gggcgaattg ggtaccgggc cccccctcga ggtcgacggt atcgataagc
     3481 ttgatatcga attcctgcag cccgggggat ccactagttc tagagcggcc gccaccgcgg
     3541 tggagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     3601 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3661 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3721 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3781 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3841 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3901 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3961 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4021 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4081 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4141 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4201 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4261 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4321 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4381 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4441 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4501 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4561 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4621 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4681 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4741 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4801 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4861 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4921 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4981 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5041 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5101 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5161 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5221 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5281 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5341 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5401 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5461 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5521 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5581 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5641 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5701 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccac
//